DE10361813A1 - Using hematopoietic growth factors for structural regeneration of tissue, useful e.g. for treating liver damage and wounds - Google Patents

Abstract

Use of hematopoietic growth factors (I), especially erythropoietin (EPO), thrombopoietin (TPO) or their derivatives, analogs and fragments, to prepare a composition for structural regeneration of tissue, is new. Independent claims are also included for the following: (1) similar use of an EPO-inducing factor (II); (2) carrier structure containing at least one (I) or (II); (3) preparing the structure of (2); and (4) composition comprising cells (A) that have been pretreated, in vitro, with (I). ACTIVITY : Hepatotropic; Osteopathic; Cardiant; Ophthalmological; Uropathic; Antiinflammatory; Antiulcer; Vulnerary; Neuroprotective. Part of the liver was removed from pigs, then the animals (a) given no treatment; (b) treated locally with 10 4> units EPO and a fibrin glue; or (c) treated systemically with 10 4> units EPO on days 0, 3, 7 and 11. After 14 days, the mean liver volume (ml) was (a) 892; (b) 894 and (c) 1073, and liver weight (g) was (a) 1001; (b) 1027 and (c) 1249; where these results for (c) are statistically significant at p less than 0.05. MECHANISM OF ACTION : Promoting regrowth of tissue.

Description

The The present invention relates to a method of inducing the structural Growth of tissue, especially in liver regeneration.

In Ontogenesis expresses growth factors that are fundamental structural and in terms of cell number numerical processes for construction of a tissue can. In the growing and in the adult organism goes the ability construct structurally and functionally intact tissues or - in the case of tissue damage - to regenerate, but largely lost. It is believed that the diminished Expression of growth factors, in turn, the expression from for control the tissue building needed for these proteins Reduction in the ability to regenerate responsible for.

It However, at least some organs are known to be themselves in the adult organism an ability to retain self-regeneration through injury processes can be induced. For example, the regeneration capacity of the liver known since ancient times. Almost all other organs can be structural Defects do not inherently bypass the original fabric restore.

The In the adult organism, the liver is usually at rest, this means non-proliferating state in which the organ is a complex variety must fulfill different metabolic functions. In vivo, however, the liver becomes by the loss of cell mass - e.g. by a liver cell damage or by a surgical procedure - again stimulated growth.

proliferating Liver tissue replaces the functional and anatomical structures However, the organ usually does not work in the desired way, but rather leads in the Usually to an enlargement and Hypertrophy of the remaining liver tissue until the original Liver cell mass is replaced. The intensity of the growth response is from the extent of Tissue loss dependent. The temporal course of liver regeneration correlates inversely proportional, that is small liver cell losses are slowly replaced, large liver cell losses much faster.

Of the Replacement of the organ mass by cell proliferation alone therefore arises no adequate basis for the therapy of a patient with significant organ damage dar are therefore different approaches for the Induction of structural growth - i. for a formative growth - of tissues but they are not yet all that satisfying Success led to have. However, such structural growth of cells would be especially for therapeutic or biotechnological processes of considerable Importance.

In The past was trying to increase the growth of cells over the Administration of growth factors, e.g. "Epidermal Growth Factor" (EGF), "Vascular Endothelial Growth Factor "(VEGF) or "Hepatocyte Growth Factor "(HGF) too induce. The effect of these factors on the proliferation of primary cells however, in vitro is limited. Their use in vivo, however, is due to their possible Side effects - e.g. the activation of oncogenes - not unproblematic.

Another approach is based on the use of complex heterologous tissue extracts, for example from the pituitary gland or the hypothalamus, for the induction of cell proliferation, for example of cultured hepatocytes (see, eg US 6,008,047 ). However, the use of animal or human tissue extracts is problematic in the context of transmissible viral diseases, such as BSE, porcine or ovine, laboratory or clinical use, and rather documents the lack of knowledge about the processes involved in building complex organ structures involved as well as the actually relevant factors and their application and effect potentials. In addition, the quality of the extracts is difficult to define, as it depends, among other things, on the source and its cultivation conditions.

Even the few findings from the classical applications of primary tissue cultures but not directly on the questions of the tissue engineering transfer. So, tissue engineering is usually ideally about patient-specific, adult cell systems that are already further differentiated as a fetal or embryonic cells. Furthermore it is the tissue engineering both in situ and in vitro to coculture situations, which in the classical application not considered become. It is on the other hand much more likely to try co-cultures Endothelial cells, macrophages and fibroblasts, as in the liver occur in the expansion of parenchymatous liver cells, since she is not wanted are.

Out WO 02/092013 A2 it is known to a patient for treatment of liver damage a therapeutically effective amount of growth hormone (Growth Hormone, GH) in order to preserve the natural regeneration capacity of the To promote liver. After that, GH has the effect of expressing the growth factor Fox M1B accelerated in hepatocytes and so liver growth to initiate again.

The However, growth hormone has a very broad and therefore non-specific Effect on the growth of tissues. Therefore, by GH gift also unwanted side effects or overreactions e.g. in shape the so-called acromegaly, d. H. an overgrowing ossification with pathological bone conditions, generated. Furthermore, it has become known in the meantime that Fox1M is upregulated in basal cell carcinoma. The Fox proteins play an important role in the regulation of growth genes in the Propagation, Differentiation and Transformation, i.a. also at the activation of so-called SONIC HEDGEHOG (Shh) signaling pathways. These in turn are involved in the activation of basal cell carcinomas in human skin. Thus, Teh et al. show (Cancer Research 2002, August 15; 62 (16): 4773-80) that the upregulation of FoxM1 in basal cell carcinomas is one of the major initiation mechanisms which makes the SONIC HEDGEHOG signaling pathways mitogenic in the exercise basal keratinocytes, causing it to the development of the widespread human carcinoma comes. This tumorigenic potential of Fox1 M agonists and the lack of specificity and ubiquitous presence In all tissues, therefore, the use of GH is promoted contrary to liver regeneration.

task It is therefore an object of the present invention to provide a method that the induction of a substantially structural growth a tissue induced. This growth should preferably become one essentially functional and structural functionality of the affected tissue.

According to the invention this Task solved through the use of hematopoietic Cytokines, their derivatives or analogues or parts for the structural and functional liver regeneration. In a particularly preferred embodiment is erythropoietin (EPO) or a derivative, parts or analog administered from it. The effect of the invention on the affected However, tissue can also be induced by the application of thrombopoietin or parts thereof.

It has surprisingly shown by the application of hematopoietic growth factors like EPO and TPO not only an increase of cells, but also a structural growth begins. This growth sets in particular on previously traumatized tissues. The growth induced thereby leads in vivo to tissue regeneration in the true sense, i. it finds not only proliferative growth, but directed, differentiated Growth to build complex structures instead.

in the Core is the use of the invention the hematopoietic Factors for the tissue regeneration essentially on two previously unknown Effects of the EPO, namely on the one hand, on the stimulation of structural growth in synchronous and coordinated form of different cell types among each other and with each other (such as fibroblasts, smooth muscle cells) Endothelial cells in the vascular area in connection with a new formation of the architecture of a complete Vessel under consideration the extracellular Matrix (collagen, elastin, fibronectin, entactin)) and a complete set of the actual parenchymatous Fabric Association. This includes e.g. the formation of hepatocytes with the connected Kupffer cells, Pit-Ito and endothelial cells (so-called non-parenchymal cells of the liver). In addition to the training of a actual vascular tree and its interconnection thus becomes a tissue regeneration according to the invention induced in the sense of restitutio ad integrum.

In the liver leads this is a mixture of sinusoidal capillaries in the final flow range and vascular incoming and outgoing Vessels next to the actual Parenchyma of hepatocytes in an orderly 3-D. Structure.

As already stated, the effect of the hematopoietic growth factors according to the invention is particularly active in traumatized tissues and cells. The term trauma is defined as opposed to the process of histogenesis (tissue formation). Thus, a trauma is a process that counteracts histogenesis as a tissue formation process in individual orga- nism at the relevant localization or nullifies the result of histogenesis. The trauma as tissue damage can be triggered by a variety of events, such as injuries, inflammation or autoimmune diseases with self-harm). These tissue damages In turn, disruption induces a variety of responses, including activation of macrophages, mast cells, and immunocompetent cells that secrete chemotactic, vasoactive, and wound-healing factors, thereby regulating systemic and regioselective mechanisms.

The Advantages of the use according to the invention the hematopoietic Growth factors, in particular the EPO, extend to the Regeneration of tissues of all four types of basic tissue, namely the Connective tissue, muscle tissue, epithelial tissue and nerve tissue. These tissues are derived ontogenetically either from the mesoderm (Connective tissue, muscle, endothelium (as a special form of the epithelium)), the endoderm (the epithelium lining the gastrointestinal tract) or the ectoderm (nerve tissue). In the past was shown that the EPO receptor both on cells of meso- and endodermal origin as well as on neuronal cells is expressed.

In leads to these tissues the use according to the invention from EPO or TPO to a local one Recruitment of the tissue-specific progenitor population (stem cells), the migration of the cells and the differentiation or transdifferentiation of cells in parenchymal and structural cells. During and before this tissue formation The cells multiply through the EPO administration.

at the application of the method according to the invention for example the liver regeneration may be a completion of the previously damaged Achieved organ to a complete parenchymatous tissue association be inclusive the formation of hepatocytes with Kupffer cells, Pit, Ito and Endothelial cells. With the continuing Training a vascular tree is thus according to the invention the tissue regeneration as restitutio ad integrum possible.

One particular advantage of the method is thus in that not only the mikokapillarisation of the regenerating tissue over a Endothelium sprouting is stimulated, but also the parenchymal regeneration and the formation of the wall structures are promoted. Only this leads to the desired Result of coordinated three-dimensional growth to construction a working one Organ.

Consequently the use according to the invention is based on an EPO effect that far exceeds the previously known EPO effect as angiogentischer factor on the Endothelial cell proliferation (Journal of Nephrology 2002 15, 97 to 103). Because microvascular Structures like capillaries and sinusoids only from endothelial cell lining exist and have no own wall structure, could starting but so far only from the angiogenic effect of the EPO about that speculation whether EPO also in neovascularization and wound healing could be of some importance (Journal of Nephrology 2002 15, 97-103). A substantiation of this speculation but so far has been out.

Therefore In the present case it is all the more important that for the first time the proof of the Effect of the EPO on synchronized and coordinated growth the vessels themselves, i.e. including the formation of wall structures and parenchymal regeneration succeeded.

One Another advantage of the method according to the invention is that structural growth is not necessarily a given organic or inorganic three-dimensional structure as a starting point needed but rather creates an organ (part) structure de novo. Consequently Can the application of hematopoietic Growth factors a significantly accelerated self-regeneration a damaged tissue induce what is in the clinical therapeutic application of the invention of great Meaning is.

In a particularly advantageous embodiment of the invention the hämtopoietic Growth factors used to regenerate a tissue induce traumatically damaged areas. In These tissue sections can thus not only a closure of the wound by the formation of a granulation tissue with incipient Angiogenesis stimulated, but the neoplasm of tissue-specific three-dimensional structure of extracellular matrix e.g. from collagen, Elastin, fibronectin or etnactin.

According to the invention hematopoietic Growth factors used. These are mainly about Thrombopoietin, erythropoietin and growth hormone (GH) as well their functionally and structurally homologous analogues, derivatives and / or Share it. In particular, however, EPO or TPO or their parts, Derivatives or analogs used. There are also their mimetic Peptides (see below). To the hematopoietic growth factors counting continue G-CSF and GM-CSF.

The hematopoietic growth factors are left quadruple helix bundle proteins oriented up-down with two overlapping loops connecting the first two and last two helix structures (Livnah O. et al., Science 1999, 283 (5404) ): 987-90 and Ultsch MH et al, Blood 1995, 86 (2): 540-7 The respective RBD domains (receptor binding domains) have a pronounced homology with EPO Thrombopoietin, erythropoietin and growth hormone bind to the MPL receptor complex. In the publication by Youssoufianh et al., Blood 1993, 819, 2223-36, Structure function and activation of the erythropoietin receptor) a productive dimerization is described. Erythropoietin and thrombopoietin, mimetic peptides (EMP and DMP) and also non-peptidic small molecules are also functional, albeit on a lower molar basis (Publications: Wrighton NC et al., Small peptides as mimetics of the protein hormone erythropoietin Science 1996, 273 (5274) 458-64 and Cwirla SE et al., Peptide agonist of the thrombopoeitin receptor as potent as the natural cytokine, Science 1997, 27653191696-9 and Qureshi SA et al., Mimicry of erythropoietin by non peptide molecules, Proceedings National Academy of Sciences PNAS USA 1999, (9621): 12156-61). The binding of EMP1 to the receptor occurs at binding sites homologous to the hot spots of growth hormone receptor compounds. The structural sequences of thrombopoietin are described in detail in FIG EP 1 201 246 A2 described. The structural sequences of erythropoietin are described in detail in the European patent application EP 84 308 654.7 described.

In a particularly advantageous embodiment of the invention, EPO is used which stimulates the formation of erythrocytes in vivo as well as the cell division and differentiation of erythrocyte progenitor cells. EPO can either be isolated from urine or produced recombinantly. The production of a recombinant human EPO is the subject of WO 86/03520. Furthermore, EPO is the subject of EP 0 148 605 . EP 0 205 564 . EP 0 209 539 . EP 0 267 678 or EP 0 411 678 ,

However, derivatives of the native or recombinant EPO can also be used. For example, an EPO derivative is known ( EP 0 640 619 B1 ), which has additional glycosylation sites and thus a higher carbohydrate content with 22 sialic acid residues. The advantage of this modification is that with an increased proportion of sisalic acid, the half-life of the EPO in the plasma increases, since only a non-sialisierte EPO can bind to the involved in EPO degradation galactose receptor of the liver. This EPO derivative - known by the abbreviation NESP (Novel Erythropoiesis Stimulating Protein or Darbepoetin) - has, in comparison to the recombinant EPO, a five-position altered amino acid sequence. However, its effect on the stimulation of the erythrocyte formation essentially corresponds to the native or recombinant EPO (Fisher, JW: Erythropoietin: Physiology and Pharmacology Update, Erythropoietin 2003, 1-14). The EP 0 640 619 B1 is fully referenced in terms of the structure and production of the NESP.

The NESP can for further extension the in vivo half life advantageously still with polyethylene glycol (PEG) are chemically conjugated without causing a change linked to biological activity would. One NESP modified in this way is known from WO 01/76640, which relates to the structure and provision of this EPO derivative in full Reference is made.

Out WO 02/49673 A2 and WO 01/02017 A2 are likewise pegylated derivatives Deri of the EPO, in addition to the prolonged plasma half-life also a higher one show clinical efficacy. For this purpose, for example, by targeted Mutagenesis 1 to 6 additional Glycosylation sites introduced into the EPO amino acid sequence. Also this derivative can be used according to the invention.

Next EPO and EPO derivatives with corresponding biological function may also other hematopoietic Growth factors such as TPO or parts thereof are used according to the invention.

at TPO (also known as c-Mpl ligand, mpl ligand, megapoietin or megakariocyte growth and development factor) around a hematopoietic Growth factor with a complex biological effect, which under the development and proliferation of megakaryocytes and others Platelets regulated. The mature TPO consists of a sequence from 332 amino acids, where the 154 amino acid N-terminal region (RBD domain) is a has substantial sequence and structural homology to EPO (20 identity and others 25% similarity). In particular, the TPO forms two highly conserved cysteine bridges, which can also be found in homologous positions in the EPO.

It has been shown in the past that the N-terminal region of the TPO is responsible for binding to the cytokine receptor and the signal cascade induced thereby via the JAK / STAT pathway (Geddis AE, Linden HM, Kaushansky K: Thrombopoietin: a pan-hematopoietic cytokine. Cytokine & Growth Factor Reviews 13 (2002) 61-73). Therefore, according to the invention, only a part of the TPO, in particular the N-terminal fragment, can also be used.

The preparation and characterization of TPO and its variants are, for example, in EP 1 201 246 , WO95 / 21919, WO95 / 21920 and WO95 / 26746. These are fully referred to for disclosure purposes. The effect of TPO on fetal hepatocytes is known from studies by E. Schmelzer (E. Schmelzer: Optimization of the culture and characterization of primary embryonic hepatocytes of the rat, Dissertation Göttingen 2002).

Suitable TPO variants are, for example, the TPO derivatives described in WO95 / 21919 or the allelic variants or species homologs described in WO95 / 21920 or WO95 / 26746 and US Pat EP 1 201 246 described pegylated TPO without being limited thereto. For the purposes of the present invention, pegylated TPO is understood as meaning TPO derivatives which are bonded to an organic polymer, such as, for example, polyethylene glycol, polypropylene glycol or polyoxyalkylene. Further variants of TPO are also understood to mean derivatives of TPO which have a sequence identity of less than 100% and nevertheless the activity of TPO, as preferably in EP 1 201 246 described own. Typically, TPO derivatives have a sequence identity of at least 70%, preferably at least 75%, more preferably at least 80%, and most preferably at least 85% compared to human TPO, including those fragments having TPO activity. A particularly preferred TPO activity in the context of the present invention is the acceleration of the proliferation, differentiation and / or maturation of megakaryocytes or megakaryocyte precursors into platelet-producing forms of these cells by TPO or variants thereof.

According to the invention, EPO and TPO in both their human and non-human form be used. For example, human TPO with pig or mouse TPO a homology of over 70%.

In an alternative embodiment the invention it is possible the patient is not hematopoietic Growth factors themselves, but to administer factors that the expression of growth factors in the traumatized tissues induce.

So is described, for example, by Naughton BA et al. (Age-related Variations in hepatic regeneration and erythropoietin production in the council, Am J Anat. 1977 Jul; 149 (3): 431-8) that erythropoietin mainly in the liver and spleen during neonatal phases is formed and that in Nephrektromie the phenomenon of hepatic EPO formation after a hepatectomy increase again can. This ability decreases with age. Later was then recognized (hepatic regeneration and erythropoietin production in the rat.Naughton BA, Kaplan SM, Roy M, Burdowski AJ, Gordon AS, Piliero SJ. Science. 1977 Apr 15; 196 (4287): 301-2. ) that one regenerating liver produces erythropoietin in response to hypoxia. It has been found that EPO production is dependent on the stage of liver regeneration, where the highest EPO Concentrations found in the phase of the strongest liver growth were.

Dornfest et al. described in 1981 (Recovery of an erythropoietin inducing factor from the regenerating rat liver. Dornfest BS, Naughton BA, Kolks GA, Liu P, Piliero SJ, Gordon AS. Ann Clin Lab Sci. 1981 Jan-Feb; 11 (1): 37-46.) That the liver is the main source for the formation of an EPO-inducing Factor is. In nephrectomy, the formation of the factor in the Liver too. Thus, the effect of the invention EPO can also indirectly be achieved by the administration of a factor affecting expression induced by the EPO.

As well For example, endogenous EPO expression can be achieved by stimulation EPO secretion through medium or long-term administration of Growth Hormone (GH) can be achieved (Sohmiya, M., Y. Kato. Effect of long-term administration of recombinant human growth hormone (rhGH) on the plasma erythropoietin (EPO) and hemoglobin levels in anaemic patients with adult GH deficieny. Clinical Endocrinology (2001) 55, 749-754).

The individual concentrations of growth factors can be found in solution at about 1 to about 100 ng / ml, preferably at about 10 to about 50 ng / ml, especially at about 10 to about 20 ng / ml. At local Coatings (topical administration) may increase the concentrations of the Growth factors, however, also amount to many times.

The hematopoietic growth factors may advantageously also be used for the cultivation of Tissue cultures are used in vitro. For this purpose, the cells are cultivated in devices and processes which are particularly suitable for the tissue, for example on a grid with a cutting mesh structure of 500 m in size, in order to allow the generation of new daughter aggregates of hepatocytes again and again. Especially in vitro, combinations of EPO with GH are advantageous.

Especially can non-contact, automatically or manually controlled pumping systems are used, the z. B. consist of piston pumps or magnetically or by compressed air compression of hoses generated, directed currents produce. In the presence of endothelial cells, it may be due to shear stress in a perfused bioreactor to a spontaneous confluence of the Endothelial cells on the surfaces the aggregates come what's for the further use may be advantageous.

For the encapsulation are suitable, known in the art, suitable Materia lien, in the For example, structured forms or spaces are integrated that enable an in situ growth structure or enlargement. Alternatively, on the capsule are dispensed with and, for example, in the presence of Endothelial cells endothelialization and thus optimal Hemocompatibility achieved become.

The gift according to the invention of EPO or TPO either alone or with bigger ones Strutkurellen defects also in connection with scaffold materials to a biological tissue replacement. These carrier materials can be used in be vorbesiedelt in vitro or extracorporeal, biologically modelable (biodegradable) and are micro- and macrostructural and biochemical ideally as similar as possible to the structure to be replaced. The biochemical proximity or identity involves a re-construction of the in vivo composition by Collagen and matrix proteins (elastin, fibronectin and all matrix components of the body in tissue-specific embossing as known).

stem Cells can from the different in the body the patient's available source: bone marrow, peripheral Blood, adipose tissue, the tissue itself, umbilical cord blood or tissue. Allogeneic stem cells can are recovered accordingly, but are with immunological disadvantages afflicted. Embryonic cells can used, but are associated with the corresponding disadvantages.

In a particularly preferred embodiment can the structured growth process of the cultured cells by one with the growth factors according to the invention coated matrix can be induced. This can be done with the matrix one or more of said growth factors sequentially be treated.

If it is a biological matrix, it is usually an implant (stent, patch or catheter), a graft (eg skin graft), a carrier material for the growth of cells, eg a so-called slow release material (eg a hydrogel on the Base of fibrin and / or polymers, such as polylactide or polyhydroxyalkanoate, and / or alginates), a bone substitute material (eg tricalcium phosphate), an allogeneic, autologous or xenogeneic acellularized or non-acellularized tissue (eg heart valve, venous valve, arterial valve, Skin, vessel, aorta, tendon, comea, cartilage, bone, trachea, nerve, miniscus, intervertebral disc, ureters, urethra, or bladder (see eg. EP 0 989 867 or EP 1 172 120 )), or a matrix (eg a laminin, collagen IV and / or Matrigel matrix) or preferably a feeder layer, such as collagen I, 3T3 and / or MRC-5 feeder layer, or a collagen flow ,

The Biological matrix advantageously becomes tissue-specific Cells, progenitor cells, Bone marrow cells, peripheral blood, adipose tissue and / or fibrous tissue, e.g. with adult progenitor cells from the bone marrow, pre-colonized. The pre-colonization leads to that the regeneration process already begins in vitro with it the implantation of the matrix in the body the regeneration time shortened in vivo is.

The used matrices can be activated before. Activation of the biological matrix or support structure For example, by plasma ionization, e.g. under use of hydrogen peroxide, or by laser activation.

alternative can be a coating with a biodegradable (bio) polymer layer, containing the one or more growth factors mentioned) become. For example, fibrin, plasma, blood, Collagen and / or polylactides.

The method according to the invention is particularly suitable for adult cells, ie primarily differentiated cells which no longer have an embryonic or fetal phenotype. It is particularly suitable for human adult cells, for example for adult progenitor cells, tissue-specific cells, preferably osteoblast th, fibroblasts, hepatocytes and / or smooth muscle cells.

Next a termination or reduction of the supply of the described growth factors to the culture are suitable for termination of the growth process of the invention also somatostatin and / or TGF beta and / or prostaglandins.

It It is particularly advantageous that methods of the invention are local in vivo use. These can be the Growth factors e.g. on the resection surface of an organ (eg Liver) are applied. You can topically or over be applied locally or systemically using a catheter. In case of liver resection can they alternatively or additionally before, while or applied after the procedure. Likewise it is possible the Growth factors (e.g., for production cartilage regeneration) directly into the affected tissue or Inject the joint. With that you can the growth factors over the synovial fluid act directly on the formation of a new cartilage structure.

In a particular embodiment can additionally to the hematopoietic Growth factors of one or more of the following growth factors be administered: "Transforming Growth Factor beta "(TGF beta), prostaglandins, granulocyte (macrophage) -stimulating Factor (G (M) -CSF), "Growth Hormone Releasing Hormones "(GHRH)," Thyrotropin-releasing Homone "(TRH)," Gonadotropin-releasing Hormone "(GnRH)," Corticotropin releasing Hormones "(CRH), Dopamine, "Antidiuretic Hormone "(ADH), oxytocin, Prolactin, adrenocorticotropin, beta-celltropin, lutroprotein and / or Vasopressin used or in addition one or more nerve regeneration factors, preferably nerve growth factor (NGF) and / or one or more multiple vascular regeneration factors, preferably "Vascular Endothelial Growth Factor "(VEGF) and / or" Plateled Derived Growth Factor "(PDGF).

The hematopoietic Growth factors parenterally as injectable microspheres, erodible particles, Polymer compounds (with polypeptide, polyglycolic acid), liposomes and small ones Particles are applied. Injectable or implantable drug Delivery devices are also possible. The substances can also be inhaled, subcutaneously, injected intramuscularly or - for the topical Application - as be given cutaneous plaster.

concomitant can Substances are mixed with the patches that reach a local hyperemia and thereby the skin permeability increase (for example bee venom). Hyperionic structures with and without direct EPO coupling or coating of the salts can barrier the skin can be transported better and can be favorably preferred ionizing structures are connected. Also negative ionization is possible. The factors can with almond oil or jojoba oil for transport improvement by mucous membranes in the intestinal area or the Skin can be used.

A Compound with polyethylene glycol (PEG) is possible to transport barriers (Mucosa in the mouth, stomach, intestine, mucous membranes, cornea) to overcome.

It It is known that prepared from proteins mixed preparations can. Suspensions, gel emulsions, solid compounds, dehydrated or lyophilized powders are possible. The growth factors can be absorbed on particles or be encapsulated.

It may be particularly advantageous stem cells (Progenitors in the actual Senses) together with EPO / TPO for tissue regeneration, in order to recruit the tissue cells from the 4 basic tissue types for the Significantly accelerate the regeneration process. EPO / TPO and the other factors mentioned mixed with stem cells and e.g. Fibrin glue as carrier matrix be applied. If necessary, the carrier matrix can be omitted or by a stronger one pre-structured and shaped carrier matrix be replaced. The factors can even without any biological carrier matrix, for example only in aqueous Suspension can be administered systemically or topically.

The gift according to the invention from EPO enhances this tissue regeneration by tissue-specific embossing the stem cells and differentiation following an increase and integration and coordinate growth in terms of the types of basic tissues.

1. Applications

1. squamous epithelia

According to the invention EPO used to promote the formation of basement membranes (extracellular support layer under epithelium). EPO supported the formation of the border area of the papillae.

EPO is used according to the invention, to the formation of glandular epithelium to support. EPO leads for regeneration plate-shaped isoprismatic and prismatic epithelia (squamous cell esophagus, high prismatic epithelium in the vas deferens, transitional epithelium of the bladder).

By EPO is undergoing re-education in the structural growth process the desmosomes, d. H. the connectors between the cells and their attachment structures. It also comes to the education of so-called Microvilli on the epithelia, for example of the small intestine (Epididymis, trachea).

EPO has a direct effect on the regeneration of the single-layered Squamous epithelium, the endothelial lining of heart, blood and lymph vessels and of the high prismatic epithelium (stomach, small and large intestine, gallbladder, Fallopian tube, uterus). EPO is used according to the invention to double row Epithelium (salivary glands the oral cavities, Nasolacrimal duct, Epididymis, spermatic duct). EPO according to the invention also for the high prismatic Epithelium (nasal cavity, Epipharinx, larynx, tracheal bronchial tree, Urethra, Eustachian tube), further to the education of not horned and horny Squamous epithelium used. The structure-promoting aspect of parenchymal regeneration supported by the EPO the formation of exocrine glands. Which includes among other things the pancreas, glands in the small intestine epithelium (Goblet cells, submandibular gland, islet cells for insulin production). EPO supported the regeneration of the endocrine glands.

2. connective tissue

EPO may be the basic components of connective tissue in coordination with the surrounding parenchymal gland structures restore. The basic ingredients that influence this are the basic substance (glucosaminoglycans), the coordination with so-called plasma cells, fat cells, blood vessels and surrounding smooth muscle cells, next to the connective tissue cells, Fibroblasts, mast cells and collagen fibers and the elastin fibers and inserted in between Macrophages (as well as Kupffer, Pit, Ito cells in the liver) and Kapillarenendothelzellen. Have capillaries, only one endothelial cell lining.

in this connection However, EPO has a broader coordinative aspect to the above Microenvironment. This becomes traumatic in one Initiator state offset. Therapeutically EPO can be given according to the invention if after traumatic injury these initiation cascades should be accelerated. EPO hereby supports according to the invention as a catalyst and systemic and topical mediator of tissue regeneration the immunomodulatory effect regionally induced by macrophages, the immunological in macrophages in conjunction with plasma cells Modulation and release of interleukins and cytokines. By this interaction competence according to the invention will the high regioselectivity the therapeutic administration of EPO / TPO at the place of need (trauma, inflammation, autoimmune disease) reached.

EPO coordinates in the therapeutic application according to the invention this regional immunostimulation with the systemic endogenous response. EPO promotes thereby the start of the inflammatory reaction in the sense of a cascade acceleration as a therapeutic effect. It increases the release of endogenous growth hormones and controlled hence its side effect potential. A systemic dose of Growth hormone is thereby avoidable and its uncontrolled Effect regarding a potential tumorogenicity avoided in the context of the physiological effect situation.

connective tissue can Restored according to the invention become. Which includes collagen fibers, reticular Fibers, for example lymph nodes, elastic fibers and training the basic substance (proteoglycans, glycosaminoglycans, hyaluronic acid, chondroitin sulphates, Dermatan sulfate, keratan sulfate, heparan sulfate, heparin). The modulated Connective tissue cells are the fibroblasts, reticular cells, in particular in lymphoid tissues and organs, mesenchymal cells, fat cells, Monocytes and macrophages. It is known that monocytes become too Can differentiate macrophages. EPO can reduce macrophages in the inflammation and in the infection reaction regarding the subsequent and necessary commercial repair and 3-D regeneration accelerate.

The Chemotactic functions of macrophages are co-coordinated by EPO. The local, monocytes located in the connective tissue can enter tissue recruiting be sucked in by EPO, causing them systemically and locally available Progenitors and for a local repair can contribute directly.

EPO coordinates the function of the retikoenterial system (RES). This becomes reticulate in a network Fibers joined together and gets the endothelial cells from liver, Spleen and bone marrow with one. In addition, by EPO, the lymphocyte activity, the Plasma cells, macrophages and - immunomodulatory - the Coordinated mast cells.

in the Connection to traumatic events EPO couples the known individual functions of these cells with structure construction and regioselective tissue regeneration. After that it comes over Rückopplungseffekte to subside the inflammatory reactions and thus also to a decay of autoimmune phenomena, that too over Antibody formation can be taught. In this case, EPO according to the invention becomes therapeutic used as a drug.

at The tissue regeneration leads to the formation of connective tissue structures and the development of loose connective tissue structures (subcutis of the skin, in hollow organs, little specialized connective tissue). EPO leads to Formation of the connective tissue (disordered and ordered connective tissue, training in bands, Ligaments, fascia, tendons and coordination with elastic Structures and their three-dimensional networking, the tissue-specific mixed or gelatinous pronounced).

EPO supports the structure of the reticulum cells, in particular the reticular connective tissue restore. EPO supported the structure of the adipose tissue and the progenitors arranged in it through recruitment and deployment of EPO propaganda functions.

By the effect of the invention it comes to improving the support functions and the power transmission these connective tissue components. It comes to improving the mass transfer, because the connective tissue carries body vessels. It comes to improving the memory functions of the connective tissue structures with an accumulation of lipids, glycosaminoglycans. EPO performs therapeutically to improve the water and electrolyte function.

It comes through EPO to improve the protective function, as it rebuild mechanical barriers Pathogenic pathogens comes. It comes to reparative processes following inflammation, as the tissue structures are restored. EPO supports according to the invention the parenchyma and structural regeneration.

This is in contrast to the formation of granulation tissues (like this one for are characterized by a so-called wound healing), in the sense a wound healing to replacement tissues and thus only to replacement functions to lead. This refers to the formation of scar tissue and loose fibrous Tissues that can achieve defect coverage, but with high functional losses are associated. In accordance with the invention EPO here a structure-forming process in the sense of the recovery of the former fabric. In particularly large defects can combinations with placeholder materials, however, the structure-building process support and promote (morphological and functional). This is very important for the tissue Engineering, since any scaffold materials used for integration (e.g. support materials for vessels, heart valves, Grafts) can be combined with EPO. Permanent implants (non-biodegradable materials) can help improve tissue integration coated (e.g., metallic and ceramic prostheses).

3. Blood vessel and lymph circulation system

EPO leads to a rebuilding of the blood vessel system and also of the large blood vessels - not only the capillary structures, as previously known. To the structures of huge Blood vessels also belong the veins and arteries (large and small systemic circulation).

Furthermore, EPO facilitates regeneration processes within tissue structures that also affect the lymphatic circulation system and the lymphatic vessels themselves. Recovery through EPO integration affects not only the tunica intima, the endothelium, but above all the tunica media, which consists of connective tissue and muscle cells. Furthermore, the Tunica Externa, which forms a longitudinal connective tissue layer. Important in the formation of the vessels are the different types of arteries. So there are arteries of the elastic muscular building type and different connective tissue structure, which are mitinduziert by regenerative processes, namely the Membrana elastica interna and the Membrana Elastica Externa, as well as the formation of vasovasal vessels. Here, fat cells are integrated into the wall layers. In elastic-type arteries, EPO facilitates the regeneration of the Membrana elastica interna in addition to the tunica externa with the vasovasols. In the case of arteries of the muscular type, cross section 0.1 to 7mm, EPO allows a strong smooth muscle formation in the media. Furthermore, EPO allows the formation of the subendothelial connective tissue layer. It comes to the restoration of typical forms of construction, such as the tunica intima, as previously known as endothelial cell component, but also especially the Membrana elastica interna, the elastic phase structures, muscle cells, Tunica Media, Membrana Elastica and Tunica Externa. Also spatial geometric arrangements such as the formation of muscular venules in the area of accompanying venules in a specific geometric arrangement of the neighborhood are made possible by the therapeutic EPO administration.

It are not 2-dimensional connection processes with one Conglomerate thrust, but to specific 3-dimensional Structure formation processes. EPO and TPO are thus according to the invention not with the known proliferation factors in the sense classical Equate growth factors, but rather act in the sense of "3-D regeneration factors".

Similar Results are found in the area of EPO stimulation in veins with the training of Tunica Intima, Tunica Media and Tunica Adventitia. Furthermore you can the venous valve structures especially in connection with scaffold Materials are regenerated into geometric shapes, or too scaffolds as far as this in larger vessel areas after traumatic injuries and degenerative processes is possible.

at larger volume defects can in certain circumstances support materials be implanted with, so that the EPO effect on the remodeling process and integration into the overall organism is focused. at complete regenerative processes, especially in parenchymatous structures However, these vessels become formed with.

It there is no restriction to different vascular structures, e.g. Arteriovenous Anastomoses. In conjunction with a therapeutically given EPO administration, in particular also age-related changes of the blood vessels and degenerative Structures are revised. In the aging process, it comes to training smooth longitudinal musculature structures. Degenerative processes mean according to the construction principle of the tissue structures Applied Neostructuring Activities. These can be as de novo produced vessels in tissue structures are constructed and are not primarily pathologically altered.

The inflammatory reactions can Modeled according to the invention by means of EPO administration become. By means of EPO administration, there are regeneration processes in the area of the endocardium as well as the myocardium. The epicard is a leaf of the Herzbeutels, which is led by EPO gift into a regeneration. The parietal leaf pericardium represents a reputable membrane that consists of a Mesothelial layer, which rests on a connective tissue layer consists. In the area of the heart, there is a regeneration of the heart valves and also the chordae tendinäe especially when structural aids (scaffolds) communicate with stem cell colonization and EPO administration are brought about. The re-education the conduction system can be done after regional destruction. Which includes also Purkinje fibers, as well as the Atrioventicular Knot, which also repairs can be. additionally become lymphatic vessels, nerves, plexuses, Corona artery structures combined with each other.

Of the Structure of the lymphatic vessels and lymphatic capillaries carried out by EPO administration in accordance with the normal form and collection vessels (Tunica Intima, Tunica Medica and Tunica Adventitia). Regenerative processes can also affect the area of the thoracic duct.

valvular and collecting containers Wrinkles, which are also integrated in structural building processes become. In the blood vessel system There is an activation of angioplasty, which in part peripheral but also regional Progenitoren be recruited. First it comes to the plant of capillaries, which consist of endothelial cell structures consist. Important in the scope of EPO, however, is that The structure structure can be initiated by the downstream muscular Structures and connective tissue structures are built.

The The conduction system plays a major role in the coordination of the EPO Effect, since there are interactions to the modulating cells and nodes gives.

To belong to the cells of the immune system Lymphocytes, macrophages, plasma cells, stem cells. The meshwork reticular Connective tissue connects the cells of the immune system in the immune system. By EPO administration leads to an accelerated repair of the reticular connective tissue, the reticulum cells and the reticulum fibers. Likewise, thymus structures be repaired again. Histologically, there are bark and marrow structures restored. Histologically, there is one in the EPO gift Regeneration also of lymph nodes, which in turn intermediate sine, Marksinus, marginal sinus, capsule and connective tissue septa, but how also laxative Lymphatic vessels, lymph nodes or Mark strands can train.

The histological organization corresponds to after the repair process and the coordinated summary of a common structure with capsule and connective tissue septa, hilus connective tissue and afferent / draining lymphatic vessels, lymph nodules, medullary tracts, and efferent Lymphatic vessels.

renewable Principles can also in the area of the spleen to restore the normal capsule, of the particle structures and the hilum. We continue to find for the Restoration of the white pulp Malpighikörperchen the spleen sinus plates in the milk strands. The blood circulation of Spleen is restored, with spleen arteries, spleen sinus veins and pulpavens together become. The white pulp and the connection of the lymphatic sheaths is restored.

By EPO administration after injury leads to a three-dimensional interaction the basal membranes, the reticulum cells, the reticulum fibers and the erythrocytes. It comes to training open and closed Circulatory systems. Here is the interaction with macrophages, the from bone marrow cells can arise, important to endogenous Cascades within the body and the mediation of growth mediators (growth hormone) to be able to.

4. skin / cutis

reparative Processes after burns, trauma, scalds, mechanical injuries or inflammation in the area of the skin are very diverse. By gift of EPO or TPO can regenerate with and without structural substitutes be achieved.

Especially in chronic diseases, circulatory disorders, diabetic ulcers, phenomena also immunological nature EPO can intervene here modeling. Histologically, there is a re-education of a normal Epidermis, dermis and a subcutaneous, which have appropriate vascularization achieved with macrostructures. On the palms and soles it comes to training of the stratum basale, stratum granulosum, stratum lucidum, stratum coneum. At the dermis and the outside it comes to training of dermal papillae. At the epidermis (inner surface) become the corresponding epidermis recesses formed again. In addition will be Appendix structures and their training from Progenitoren supported. One The problem with previous tissue engineering was the connection of structural Repair with training of skin appendages in the area of the skin.

By Gift of EPO Interaction processes in the field of the human melanocyte system be achieved. Physiological feedback processes between Eyes, meninges and the formation of pigmentation are considered, since it is not isolated cell connections, but organzypische Regeneration products is.

reparative Processes of dermal involvement include muscular structures (musculus arector pili), tactile structures, outer root sheath the hair and inner root sheath and also the hair bulb structure. This may be outstanding into the fatty tissue. When the hair and hair follicles comes It contributes to the formation of connective tissue root sheaths, which are an important Function structure in the regeneration includes. Make it glass skin, outer root sheath, inner root sheath and the final hair. Similar Processes can be supported and brought about in the field of nail regeneration. Within the tissue structures, sweat glands are also formed. In the Skin structures become perifollicular network structures, networks, subcapillary arterial networks, term Venetian nets again trained.

5. digestive system

The EPO administration restores the innermost layer of the mucous membrane, tunica mucosa and connective tissue layer of the so-called lamina propria. The deepest layer forms a layer of smooth muscle the lamina muscularis mucosae. The layer structure is as follows: serosa cells then muscularis, Submucosa and mucosa. These structures are permeated by digestive glands and mucosal glands are located in the mucosa. In addition, submucosal glands and the plexus myintericus (Auerbach) and the plexus submucosa (Meissner) regenerate. In the area of the tongue, the leaf-shaped papillae regenerate. In the area of the mouth, taste buds are also regenerated in the area of tissue regeneration. These are composed of basal cell type 4, support cell type 1, receptor cell type 3, glycoproteins and taste spores. The seromucous salivary gland consists of striated pieces, shunts, mucous tails, basement membranes, myoepithelial basket cells and serous endings. Important reparative processes in the case of paradontosis can also occur in the area of the tooth area. For example, this relates to the structures of the periodontium in conjunction with the alveolar bone and the root canal with apex. The reparative processes can be accomplished by adding stem cell-populated calcium phosphate structures or other mineral replacement materials coated with EPO. The EPO-sponsored training in a regeneration process occurs in the area of cysts in the region of the maxillary sinus, mandibular structures, after root removal and repair of these structures on-site in vivo.

The appropriate materials and odontoblasts can be carried out in vitro as well EPO gift to be prepared.

osteochondral Structures such as the osteochondral cylinder can be found in Prepared in vitro and vivo with EPO administration implanted growth-promoting become. The formation of root canal structures with neoinnervation will over EPO coated lead structures infused are promoted. Here help molecules the extracellular Matrix, which in coating processes also called peptide structures brought can be. The enamel organ structures together with pulpodontoplasts, intermediate zone melt pulp can supported in the growth process after injuries and traumatic processes. The reparative and regenerative processes focus on the area of the surrounding connective tissue and the integration of the Tooth structure or an implant in the area of the jaw. The pulpal connective tissue and training with blood vessels, the In and out of the apex of the root canal plays an important role here for the Innervation. The peridontium (periodontal skin) is regenerated in this way. The Sharpey fibers run through the periodontal membrane and are in the cement or Anchored alveolar bone.

EPO leads to regenerative processes in this area after inflammation and traumatic reactions. In the area of the gums comes it by EPO administration in inflammatory phenomena Paradontosis to improve the reparative process.

Maw:

renewable Processes can also by EPO administration on the pharyngeal area in the recovery of the plasmatic multi-row epithelium. Here too can Tunica muscularis and Tunica adventitia coordinated restored become. In the lymphocyte structures within the palatine tonsil Kryptastrukturen and capsule structures are restored. Regenerative processes can also inflamed Structures, as in tonsillitis regioselective in regeneration promote.

Hull / intestine:

in the Intestinal area, there are highly regenerative epithelial portions that are within can renew from 24 to 48 hours. In inflammatory reactions it comes to a disturbance this reparative activity, so supportive Effects by EPO / TPO allow here a compensation reaction of the body can. hereby it comes to the re-education of epithelial structures such as lamina propria, Tunica mucosa; Lamina muscularis mucosae, tunica submuscosa, ring musculature, Tunica muscolaris and longitudinal muscles and Tunica adventitia, which as reparative processes the entire Would pass through the intestinal wall area, so that a selective, superficial also intramural and deepmural regeneration is possible. These principles can be over the entire area of the gastrointestinal tract starting from the pharynx, mouth into the esophagus and continued into the rectum become. In the esophagus can the lamina muscularis mucosae, lamina propria and the multilayered Squamous epithelium be regenerated. Serous structures are integrated and repaired by EPO gift.

Reconstruction of the gastric wall in particular the area of the gastric areas with gastric pits, gastric glands, lamina muscularis mucosae, mesothelium on the outside, then subserous connective tissue, tunica muscolaris, tunica submucosa and tunica mucosae is possible. Parietal cells, gastric glands, main cells, lamina propria, muscularis mucosae and tunica mucosae are regenerated in the structures, as is the surface epithelium. Regenerative processes also in the area of the cells of the corpus fundus gland preparation There is a rebuilding of main cells. The connection to parietal cells is given. The stimulation achieved by EPO according to the invention also involves the regeneration of cells which release vasoactive substances such as polypeptides and the formation of enterochromaffin cells (ECN), which release serotonin. Gastric glands can be regenerated. It is known that the gastric surface epithelium can regenerate within three days. In case of severe traumatic, inflammatory injuries, regeneration disorders, the administration of EPO helps to restore this regenerative ability. In the area of the small intestine, the lamina propria, the lamina muscularis mucosa, the solitary follicles, the tunica submucosa, the musculature of the ring, the longitudinal musculature, the tunica serosa regenerate. In the structures, the villi are re-educated, as well as the solitary follicle and Kerkring's folds. Likewise, the crypts are restored. The crypts contain so-called Paneth cells. These can form secretion generator in the glands. Peyer's Plaque is located in the lamina propria of the duodenum. The end pieces of Brunner's glands exhibit characteristic mucosal cells after EPO regeneration, a bright cytoplasm and flat cell nuclei lying at the cell base. The colon regeneration in EPO administration plays an important role especially after surgical distances.

6. Skeleton

EPO leads to Regeneration of the skeletal tissue with the division into tight collagenous connective tissue, cartilage and bone tissue. Both Promotes chondrocytes EPO the formation of intercellular substances and leads to functional Improvements, the chondrocyte structures. EPO supports the Formation of the growth zones and the formation of the pericondium (surrounding connective tissue structure, exception joint) after traumatic Effects. In adults can As is known, cartilage cells no longer multiply as the cartilage-forming activity of the pericondium on the time of body growth limited to adulthood is. By gift of EPO can therapeutically influences these regeneration processes and the otherwise occurring formation of connective tissue cells as defect filling material avoided become. Partially can hereby the plastic process, i. the reshuffling of fibroblasts and chondrocytes play a role in the recruitment of the EPO. Therapeutically EPO is used in the regeneration of cartilage structures, such as. Hyaline cartilage in the area of the joint surfaces for the laryngeal ends, Windpipe, Bronchi and part of the nasal skeleton. According to the invention promotes EPO the formation of cartilage capsules and the formation of proteoglycans / collagens. According to the invention is a Connection with stem cells possible. But even isolated EPO is used to elastic cartilage too regenerate, e.g. localized in the epiglottis cartilage area is. Which includes also the cartilage of the outer ear and the Eustachian tube. EPO is used according to the invention to also fibrocartilage restore in the joint area of joints. These include the Regeneration processes in the area of the intervertebral discs, intraarticular processes, discs and menisci. According to the invention can in adult organism the otherwise lost regenerative changes of the cartilage to be reactivated, regeneration, rebuilding and start the proteoglycan / collagen synthesis of the chondrocytes again. The calcification of cartilage may be undesirable Preventing posts.

According to the invention leads EPO in therapeutic areas for the coordinated 3-D regeneration of bone tissue as Complete structure including the vessels. at bigger defects can Replacement structures of inorganic or biological support materials introduced become. Here can be a combination of phase-pure tricalcium phosphate and an EPO coating.

alternative can Hydroxyl appatites or even biological bone allogeneic or xenogeneic origin after appropriate treatment accordingly be used. Acts according to the invention the gift of EPO, on the periosteum and the endost and leads to one Recruitment and propagation of the progenitors located there. After that it comes to the formation of lamellae, formation of osteocytes and the corresponding lacunae and canaliculi. According to the invention EPO used for bone regeneration, and it comes to training the Haverskanäle, canaliculi the lamellae, the lacunae, concentric lamellae, the one adjacent slat system boundary and connection allow. Surprisingly become Volkmann channels as bony Streets for the blood vessels found. EPO stimulates osteoprogenitor induction, osteoblasts, osteocytes and osteoclasts. EPO leads in osteoblasts to a neosynthesis and induction of intercellular substances of the bone. Tissue formation involves the formation of bone-specific Cells. EPO promotes the training of ossification channels and the training of osseous and the integration of vascular Connective tissue. Starting from injuries, e.g. Accidents, trauma but also inflammatory reactions, can EPO the coordination tasks in restoring the Take over fabric structures, so that zones of cartilage degradation, bone regeneration with zones of Cartilage calcification, zones of chondrocyte hypertrophy, division zones (Columns, cartilage and reserve cartilage zones (high cartilage)) arise.

According to the invention promotes EPO the vasculogenesis also in the area of the bone, in particular the training an arteria nutricia, the metaphyseal vessels and the epiphyseal vessels. In addition will be the periost vessels site-specific integrated.

EPO promotes the formation of the nucleus pulposus and annulus pulposus as well as the hyaline cartilage in the area of the intervertebral discs. In joint structures become outer ligaments, Inner ligaments and capsule reinforcement bands in the Education encouraged. Repair processes of the synovial membrane are included here. According to the invention promotes EPO after inflammatory reactions the re-education of smooth muscles, toned muscles and myocardial tissue.

7. Smooth musculature

The Histogenesis of smooth muscle cells is under development by EPO and differentiation promoted to myoblasts from mesenchymal cells. In the skeletal muscle promotes EPO the formation of the perimysium and the endomysium. By EPO it comes to an integration and coordination of the ingrowth of motor Nerve fibers and the formation of motor end plates. EPO promotes the Histogenesis of the heart muscle. The heart muscle usually has no or very little regeneration capacity. EPO is used according to the invention to the tissue regeneration of smooth muscle cell structures, the Achieve skeletal muscle and cardiac muscle tissue and a to achieve imunmodulatory decay effect by feedback. This is important for the therapy with inflammatory autoimmune muscle diseases.

8. Nerve tissue

In The three-dimensional tissue regeneration plays the connection with Nervous systems and innovation processes play a major role. According to the invention EPO to coordinate vasculogenesis with neurogenesis and the merge of these structures in the innovation of parenchymal glands, tissues and organs used. This leads to the re-education of synaptic contacts, the formation of neurons as laminas and vertical Units and the training of nerve fibers and fiber tracts. Connecting structures in the area of the ganglia plexes are caused. The cranial nerves and cranial nerves are involved, the spinal nerves are regenerated after separations and both efferent motor as well as afferent or sensitive structures are switched again. Supports can nerve guide made of biopolymers, (biodegradable or non-biodegradable (Silicone, polyurethane)) or of biological origin, or of synthetic origin (e.g., collagen, elastins, fibronectins, Polylactide, polyhydroxybutyrate, silk) and in combination with stem cells can be used. The stem cells can be of mesenchymal origin. You can also choose from the fabric structures the neural structures are recruited directly, and it is not is an apoptosis prevention, but an initiation of Neurite growth and multiplication in a directional three-dimensional Structure.

An important effect of the EPO effect is the tissue regeneration in the connective tissue structures of the central nervous system, especially the meninges and their accompanying blood vessels. This effect is very important for the regeneration of ganglia and sensory organs. According to the invention, the formation of axon structures and the formation of axons is made possible. Electrophysiological activity and axonal transport is restored once cross-linking is achieved. By combined administration with vitamins, for example vitamin C, these processes can be supported. However, the effect according to the invention in tissue regeneration in the interaction between activation by local plasma cells, macrophages, lymphocytes may be important for mast cells which lead to cellular recruitment and reparative behavior (after therapeutic EPO administration). In the central nervous system, EPO according to the invention can also be used for regeneration in the area of degenerate structures. These include Parkinson's disease and degeneration in the striatum area, as well as Alzheimer's disease and amyotrophic lateral sclerosis, which also radiates to the periphery, as well as multiple sclerosis and organically induced depression. Important here is the treatment with inflammatory stimuli, so that the activation cascade, which is determined by EPO, can be initiated. A stereotactic implantation with mesenchymal progenitors and neuronal progenitor cells as well as the use of local progenitons of regional and peripheral monocytes can be therapeutically enhanced by EPO and led to regeneration. The goal is the formation of new neuronal cells of the central nervous system. According to the invention, there is then a functional uptake of the cells with the formation of neuroactive peptides and endorphins and, for example, serotonin, dopamine, norepinephrine, actylcholine. Synaptic processes are reconnected. In accompanying functions of regeneration also not the neural cells of the CNS, such as Examples include the protoplasmic astrocytes, the fibrillar astrocyte, oligodendrocytes, microlia (glioblasts) and the ependyma. The nerve tissue is formed by EPO not only in a neuronal regeneration, but also in the accompanying supporting cell structure of the so-called neuroglia. In proportionality, the neuroglial cells surpass the neuronal cells by up to a factor of 10. They form the connective tissue skeleton of the CNS and guide the blood vessels. This has an important regenerative component for structural reconstruction. Ependyma is the epithelial lining of the inner surfaces of the ventricles in the CNS.

EPO leads according to the invention to a Regeneration of the axon sheaths, a so-called Schwannschenscheide. EPO promotes the formation of myolin vagina (Myelin sheath). In the CNS, oligodendrocytes provide myolin sheaths. This is therapeutically particularly important in diseases of the sheath formation. at the peripheral nerves are induced by the accompanying cells, ganglia to change clothes. The Schwann cells are stimulated by EPO in the area the regeneration process the new formation of the peripheral nerves to allow with. It comes here to the formation of interperiodic lines of the myelin segments.

EPO promotes According to the invention the training the gray and white Substance and its tissue-specific differentiation in the CNS. EPO supports the formation of ganglia (accumulation of nerve cells outside of the CNS). After traumatic situations it comes to regeneration of these structures. The regeneration processes also in regenerative Area will be over EPO modulation led to neoinnervation, starting from Lacrimal gland, mandibular gland, subjacent gland as well Parotid gland, Heart, larynx, trachea, Bronchi, lungs, stomach, small intestine, Blood vessels of the abdomen and other blood vessels, liver, Gallbladder, bile ducts, Pancreas, Adrenal medulla, large intestine, rectum, kidney, urinary bladder and genital organs Neoinnervation comes as a reparative process. In the area sympathetic frontier cord and sympathetic ganglion (Ramus communicans albus and ramuns communicans griseus and nervus splancnicus, Ganglium celiacum and ganglium mesintericum superius) to reparative processes. The nerve endings can be through the regeneration processes also integrated into the multi-layered squamous epithelial structures become.

histological become end-piston structures, corpuscles and so-called Pacini bodies restored to the sensitive structures. EPO supports the Formation of the dura mater, whose structure again connective tissue leitstruktu reindeer contains sensitive nervous blood vessels. Thereby it comes to the training subdural space (training of the duramatis spinalis) and the epidural space. The dura mater encephalica is liable at the skull bone and is due to the EPO stimulation as part of the regeneration process rebuilt. The arachnoidia extends into the subarachnoid space. EPO regeneration processes also stimulate recovery of the subarachnoid space and the soft meninges. This can relationships to the cardiovascular Record (Virchow space). EPO leads to restore the thin connective tissue layer the the surface of the brain and spinal cord covered. It comes to the regeneration of the venous sinuses of the arachnoidiazots. In particular, the plexus chorioidius and the brain ventricles become regenerated. The gift according to the invention of EPO leads in inflammatory processes to a re-education of the blood-brain barrier by structural Tissue regeneration. The functional significance of this regeneration which are over the entire brain tissue including the medulla oblongata with the exception of the neurohypophysis, is of great importance for the homeostasis of the CNS. The barrier is restored with astrocytes by EPO administration re-educated.

histogenesis of the nervous system by EPO exposure

After trauma in the central nervous area and degenerative processes, the interaction between glial cells, oligodendrocytes, neurons, subdural space and blood vessels is disturbed. These spatial connections and the accompanying cells are physiologically involved in a histogenesis. Nevertheless, it comes to healing problems in injury, since the neuronal adult tissues are not capable of regenerating to the desired extent. By administering EPO, according to the invention, this process can be accelerated in a regenerative direction and the structural restoration can be achieved by combining the structural components with the parenchyma components. In embryonic development, this developmental step follows a set schedule. EPO is a mediating structure in the CNS in connection with, for example, the nerve growth factor (NGF). The lack of regeneration capacity of the CNS in the adult organism is compensated by EPO administration in that a Recruitment of the local cells, immune modulating cells, clearance cells (macrophages) and stem cells can be done. Stem cells (eg autologous from peripheral blood CD45 positive cells, mesenchymal stem cells, from adipose tissue, from umbilical cord tissue) and other stem cells can be combined. The use of autologous tissues is preferable to the use of allogeneic cells (eg, embryonic cells) from the EPO effect in but not to be excluded).

In Combination with stereotactic surgery or infusions may be the Progenitors are site-specifically positioned. Severed or injured peripheral nerve fibers may be linked by EPO administration with lead structures, guard rails or even in conjunction with biological Structures on site (from the immediate area) in the desired Be directed area z.b. also by fibrin structures. fibroblasts structures can also be integrated in other areas with stem cells and EPO. Alternatively you can also biological matrix molecules based on collagens or elastins in recombinant form or consisting of peptide sequences, are introduced into mixtures, whereby it comes to injection solutions, by EPO gift can be taught. At the same time, the new generation can also be systemically supported by gift of EPO. Without the gift of EPO and stem cell support comes it became known as the so-called Wallerian Degeneration, which has been known since 1850 is. Along the so-called Büngner Band it comes to regeneration processes. In the CNS it comes after 12 to 24 hours after a lesion terminal degeneration phenomena. The cell body dissolves by chromolysis. The regenerative processes by EPO be accelerated lead to an axon structure. By EPO the nettle substance and the Golgi apparatus renewed. Take the swelling of the cell body while off in a physiological manner in the first week after a lesion regeneration phenomena at peripheral axons occur. By the gift of the invention from EPO it comes to accelerations of regeneration, so below one week already regeneration phenomena, like the training of philopodia and protoplasmic growth lobes. The axonal Philopodien need usually two weeks to caves in the Schwann cells to wax the distal segment. Through biological structures and Collagens can these structures are bridged during complete transection. An application form for the Nerve splint, e.g. EPO coated collagen tubes. Other substrates Like polylactide or Poldhydroxyalkanoate can also be used. The combination with stem cells and autologous or allogeneic fibrin as a lead structure is possible.

at Polymyolitis states By EPO administration a neosynthesis of the perineurium is achieved. This promotes the formation of collateral budding in the CNS.

9. liver

To viral infections of the liver (e.g., hepatitis A, B, C, D) may be come to regeneration defects of the liver. The liver cirrhosis presents the end stage of a liver regeneration defect at which it is came to connective tissue replacement of the parenchymatous cells, since the body's own Regeneration not commensurate with the rate of noxious damage can keep up with continuous regeneration.

To Trauma and surgery, e.g. in tumor resections of the Liver, there is an acute loss of viable tissue and Tissue destruction.

According to the invention comes it after administration of EPO for accelerated regeneration of hepatocytes and non-parenchymal cells of the liver to the Restitutio ad Integrum. Characteristic of this is the re-education of a normal liver lobule architecture, re-education of the interlobular Connective tissue, the central veins and the glisson triad: There will be recognizable bile ducts, Lymphatic vessels, periportal Tissue, arteries and interlobular vein adjacent to liver cells restored.

The specific formation of the intralobular reticulum fiber braid and the liver sinusoid is restored. The histological Structure after regeneration shows the typical beam structure. The Fenestrations in the area of the endothelial cells indicate a typical liver expression out. Important initiator cells after trauma are the Kupffer cells release the IL-6, IL-1, TNF-alpha.

The Gift of EPO leads to much higher Regeneration processes even ontogenetically compared to liver tissue young livers. In vitro factor increase by a factor of 10 to 30 reached.

10. Gallbladder:

In inflammation of the gallbladder and also after mechanical injuries of the gallbladder, therapeutic EPO administration restores the normal structure of the wall as follows: connective tissue (capsule, fused with liver capsule), tunica muscularis, luschka ducts, lamina propria (richly vascularized) and Tunica mucosa (with epithelium, single-layer, high prismatic). Lymphatic vessels, nerves from the area of the vagus are reconnected.

11. Pancreatic regeneration:

The EPO gift supported a regeneration of the B cells, D cells and A cells of the pancreas. The regeneration begins from the small epithelial strands that emerge from the excretory ducts. The veins and arteries of the pancreas are mitregeneriert.

The Regenerative processes affect the endocrine and exocrine parts after trauma, inflammatory Diseases or immunological autoimmune processes.

12. Nasal cavity and Paranasal sinuses:

septum nasi, vestibulum nasi, nasal cavity, Nasal turbinates are subject to regeneration after injury and EPO administration Processes. The epithelium is restored and also the lamina propria (tight, collagenous connective tissue with tubuloalveolar mucosal glands), vessels with arteriovenous anastomoses and erectile tissue, adrenergic and cholinergic nerves. In the area of the olfactory mucosa the very high epithelium and the serous bowman glands are rebuilt.

13. Larynx area:

The multi-row, highly prismatic ciliated epithelium, the lamina propria, the serous glands and the mixed - seromukösen glands become restored by EPO / TPO administration with and without stem cell combination.

14. Trachea:

Restoration typical wall structure with perichondrium, hyaline cartilage, Perichondrium, lamina propria with seromucous glands, submucosa, lamina propria and epithelium. The adventitia is regenerated on the periphery and the cartilage clips connected by smooth muscles. In the epi-hello There are goblet cells, basal cells and ciliated cells. As well endocrine (pa-) cells, type I and type II brush cells.

15th lungs:

The complex 3-D architecture and structure of the lung according to the invention as follows restored. Bronchial artery, glands, bronchus, branches of the arteria pulmonallis, bronchioles, smooth muscles and branches of the vena pulmonalis, ductus alveolari, alveoli sacs and the pleural sheath is merged in three dimensions. In the bronchopulmonary Segments become intersegmental septa, segement veins and segmental arteries regenerated. In the bronchi become epithelium, lamina propria, tunica muscularis, glands and cartilage regenerates. In the alveoli, the blood-air barrier function restored. Elastin structures are regenerated. alveolar epithelial cells (Type I and II) occur again. Blood vessels, lymphatics and nerves (pulmonary plexus) are three-dimensionally coordinated by EPO and / or stem cell integration.

16. Urinary System:

The regenerative effects in the kidney area have remained unknown until now, since EPO in chronically insufficient kidneys is substitutive for the EPO Deficiency was set. For traumatic injuries, inflammation and toxic or immunological damage occurs according to the invention EPO administration with and without stem cells / progenitors for regeneration of urological systems. In the kidney it comes to regeneration bark structure, renal calyces, medulla, medullary rays, Kidney pyramids and Columna renalis.

It comes to the regeneration of the mesangial matrix, the mesangial cells, of the glomerulus (with lamina rara interna, lamina densa, lamina rara externa. The structural relationships of collagen filament bundles, the Mesangial cells, podocytes and endothelial cells are restored. In the renal cortex become collecting tube, main piece, middle piece, macula densa, juxtaglomular Regenerates cells and Vas afferens. In the kidney's marrow comes for the re-education of the characteristic interstitial cells and the ductus papillaris.

The Urinary structure is transformed into tunica adventitia, ring musculature and longitudinal muscles the tunica muscularis, lamina propria, and epithelium (tunica mucosa) regenerated. The bladder is used to restore the epithelium, lamina propria (tunica mucosa), tunica submucosa (longitudinal and Ring muscles) and outer longitudinal muscles guided.

The female urethra is using EPO and / or stem cells to regenerate the tunica muscularis, of the venous Braid, the glandular lacunae, the lamina propra and most of it ungerminated squamous epithelium. In the male urethra becomes the pars prostatica, the musculus sphincter uretrhae and the diaphragm urogenital, utriculus prostaticus, pars membranacea, Pars spongiosa and the fossa navicularis regenerated. On the mucous membrane the pars spongiosa is followed by the corpus spongiosum. After traumatic Injuries or surgical interventions may be associated with EPO Fibrin glue with and without stem cells in the structural defects be injected or applied topically become.

17. Endocrine glands:

in the Pituitary gland, epiphysis, thyroid, parathyroid gland, pancreas (Langerhans Islands), Adrenal glands, ovaries and testicles may be traumatized by trauma, Surgery (e.g., tumor resection) can lead to tissue injury. In these cases EPO according to the invention in conjunction with stem cells from autologous bone marrow, umbilical cord stem cells, Progenitors from the peripheral blood (monocytes) or by recruiting ortständiger Progenitors lead to or support tissue regeneration. In the adenohypophysis become the basophilic cells, the arteriae hypophysales superiores and inferiores, the gomitoli and the portal vessels restored. The neurohypophysis is in its composition with pituicytes, unmarked nerve fibers from neurosycretary neurons in the hypothalamus regenerated. For this purpose, according to the invention EPO fibrin strands as Lead structures with stem cells placed between the regions. capsular structures are regenerated as in other brain regions. At the epiphysis becomes Habenula, Recessus pinealis, lobule structures, brain sands, Connective tissue septa and pia mater (capsule) restored. Nerve fibers, Pinealocytes, Astreozyten become into the specific structures connected to the epiphysis.

The thyroid is subject to frequent surgical interventions. The follicular epithelial cells, C cells, connective tissue structures are regenerated accordingly. Adrenal glands exist after regeneration according to the invention from the capsule, glomerulosa zona, zona fasciculata, zona reticularis. Spongiocytes are located in the zona fasciculata.

18. Reproductive system:

renewable Repair processes can according to the invention structural processes in ovaries to restore the functional structure lead. These are in particular the regeneration process of the Stroma ovarii, the tunica albuginea, the granulosa cells, theca folluculi interna and externa involved. The mesothelium lining will after injury closed again. In the fallopian tubes, the wall structure is restored (Epithelium, lamina propria (mucosa)), ring musculature, longitudinal muscle (muscle layer), subserosal Connective tissue, mesothelium (serosa)).

In The ampoule of the fallopian tube becomes glandular cells, lamina propria and Ciliated cells regenerated. Corpus uteri is in endometrium, myometrium and perimetrium structurally regenerated. Mucous membrane (epithelium, lamina propria), muscle layer (longitudinal muscle, musculus bulbospongiosus) and adventitia of the vagina are regenerated. Lymphatic vessels and nerve supply will be restored. In addition to the entire tissue area of the female Sex organs are also in the male genitals Repair processes after trauma and other injuries possible. So be u.a. Testes, epididymis, penis (including corpus cavernosum), Ductus seminiferus, prostate and their structural involvement in the surrounding soft and Hard tissue coordinates regenerable. The blood-supplying structures, Lymphatic vessels and nerves become reintegrated.

19. mammary gland:

Of the structured construction in lobules, Connective tissue, Milk sachets, Excretory ducts with the histological detailed structure alveoli, intra- and interlobular connective tissue, Milk passage, Myoepithelzellen takes place through regeneration again. Especially after tumor resections or plastic surgery can the chest tissue in conjunction with biologically modelable scaffolds, EP and stem cells to be rebuilt.

20. Central nervous system:

in the Brain and spinal cord area play in particular mechanical and ischemic insults with following Tissue degeneration or structural breaks a clinically significant Role. Gray and white Substance, cerebellum, midbrain and cerebrum, nuclear regions and fibrous pathways are structurally merged and regenerates.

To the Zwischenhirn counts the epithalamus, thalamus dorsalis, subthalamus and hypothalamus. The brain area comprises about 50 fields in cytoarchitecture which can be structurally regenerated. The main areas are Frontal lobes, temporal lobes, parietal lobes and occipital lobes. The septal regions, the olfactory bulb and the cortex areas can be regenerated. To the regeneration area of the basal ganglia belong the corpus striatum with the caudate nucleus and putamen, the ventral one Striatum, as well as nucleus accumbens and the nucleus basalis. By the regeneration processes can the failures motor Areas with the associated paralysis and the failures sensory neurons are revised. The damage in the field of motor Bark (e.g., by birth trauma and accidents) with the resulting spastic paralysis can be revised. At the damages the frontal horn with the accompanying flaccid paralyzes can regenerative processes can also be induced. Disorders of Hippocampal functions cause severe disturbances in the area of orientation and memory (Korsakoff syndrome). Damage to cortical neurons The associated risk of Alzheimer's disease can be contained. In cerebrovascular Insufficiency to cerebral infarction can be due to reparative processes Regeneration be initiated.

21. Eye:

The Regeneration processes extend to the cornea, conjunctiva, Iris, Schlemm's canal, ciliary muscle, lens, zonula fibers, Ora ser rata, medial rectus muscle, vitreous body, retina, vessels, dura mater, arachnoid, optic nerve, arteria and vena central retinae, Retina, choroid and dermis. The specific structure of the cornea with collagen lamella, fibroblasts, elastin fibers, collagen lamellae, bowman Membrane, epithelial layer is restored. Sclera (dermis) is regenerated in lamina fusca, substantia propria and episklera. Lens epithelia, eyelids with epidermal and dermal components, subcutaneous tissue, Sweat glands, Hair follicles, epithelium, lamina propria, tarsus, melborn gland, minor gland, musculus levator palpebrae, musculus orbicularis oculi is restored.

22nd ear:

Auricle, Middle ear, eardrum, tympanic cavity, ossicles, Musculus stapedius, musculus tensor tympani, antrum mastoideum, cellulae mastoideae and Eustachian tube, inner ear and bony Labyrinth, cochlea with associated nerves, labyrinth and Corresponding to the Corti organ are according to the invention in the involving regenerative processes.

II. Embodiments

1. Cell transplantation

hepatocytes:

liver cells become by collagenase digestion in the usual way from non-transplantable Organs or liver resectants isolated. (Bader, A., Rinkes, I.H.B., Closs, I.E., Ryan, C.M., Toner, M., Cunningham, J.M., Tompkins, G.R., Yarmush, M.L. (1992) A stable long-term hepatocyte culture System for studies of physiologic processes: Cytokine stimulation of acute phase response in rat and human hepatocytes. Biotechnol Prog. 8, 219-225.) The isolated or pre-cultured cells are in liquid Stored nitrogen. After thawing the cells according to known Protocols (Karim, N., Allmeling, C., Hengstler, J.-G., Haverich, A., Bader, A. (2000) Diazepam metabolism and albumin secretion of porcine hepatocytes in collagen sandwich after cryopreservation. Biotechnol Letters 22: 1647-1652) are the hepatocyte suspension / cultures with 100-150 IU / kg / body weight epoetin alfa (relative to the recipient). Epoetin alfa is administered in a sterile solution in a volume of 1-1.5 ml of a 10 ml hepatocyte suspension with a concentration of 2-10 Million / ml cells added.

These Suspension is injected slowly (1 ml / minute) intraportally. The Suspension may additionally with 1000 IU of heparin to avoid thrombosis.

alternative The cell / EPO mixture can also be taken under the liver capsule or directly injected into the liver parenchyma at multiple sites. For this it is recommended that the concentration of hepatocytes by the factor 2-5 increase and reduce the injected volume accordingly.

The branch channels be with commercial Fibrin glue (Baxter Tissucol) closed. Alternatively, a Tamponade be used with collagen fleece. The tamponade can as well with the epoetin alfa solution be offset. It is important to note that the tamponade the adhesion site still dry.

The fibrin glue represents a 2-component mix. Usually one component consists of fibrinogen and the other one of an activation solution with Ca ⁺⁺ and proteinase inhibitors (eg aprotinin). Epoetin alfa may be added to the activation solution by adding it to a final concentration of 100-150 IU / kg / body weight.

In analogous way Stem cells from the bone marrow, the adipose tissue, a specific parenchyma or the blood (purification from Buffy Coats, CD 34 positive cells) from umbilical cord blood and the mesenchymal cells from the Nableschnurgewebe be used.

Parallel for cell transplantation in ischemic, toxic, infectious or mechanically (traumatically) damaged areas can Cells are introduced into a fibrin glue or autologous plasma and with the addition of epoetin alfa (100-150 IU / kg / KGW) brought to polymerization become.

Parallel an EPO dose of 10000 IU s.c. to be started so that a total of 40,000 IU are applied within a week.

The The result is a factor of 2-3 increased tissue regeneration, the leads to a structural repair.

2. Postoperative gift

To extensive operations in the cardiothoracic vascular area, the visceral surgery, plastic surgery but also after extensive trauma Defective situations that can lead to a vital threat to the patient.

epoetin Alfa is given here at a concentration of 100-150 IU / kg / body weight.

By the EPO administration leads to an endogenous increase in growth hormone the factor 2, causing tissue regeneration after surgery is accelerated. The restitutio ad integrum occurs about 1-2 weeks earlier as a patient treated with comparatively no EPO administration.

EPO can also after a split liver transplant or liver transplantation be given to induce liver regeneration.

To a liver transplant, it happens that the transplanted Tissue - or with living donors - the remaining tissues are not fast enough and in sufficient quantity available as active tissue stands. In this case can already be 24 h before surgery and for Time of operation and then at 24 h intervals 100-150 IU / kg / kg Epoetin be given alfa. This leads to a significantly accelerated Liver regeneration wherein postoperatively, especially on 4-5 days one Volume increase in the ultrasound can be diagnosed.

3. Gift to operative liver resection in benign and malignant tumors

at There is a need for an extensive liver resection, an accelerated one To achieve regeneration, since the availability of a sufficient Cell mass important for the survival the patient is. After surgical resection may be at systemic 100-150 IU / kg / kg Epoetin alfa or equivalent in case of topical application (in fibrin glue, plasma).

at Suspected persistent tumor invasion or non-resectable Tumor metastasis can both systemic as well as topical according to the tumor type and the prognosis usual cytostatic drugs administered in combination with EPO.

To Resection and administration of EPO results in accelerated growth in size of the treated group by 30% relative to an untreated one Group. Especially the hepatocytes on the resection surfaces occur reinforced into the structural growth process. It comes to a complete education of the vascular tree and the fabric around it. Here are the hepatocytes in a typical manner similar to the normal organ in Lobuli Vasculature, Cell plates with nonparenchymal cells (Kupffer, Pit, Ito - and endothelial cells) arranged.

It comes to a systemic size growth. The growth in size ends when the initial size is reached.

The Group with perioperative EPO administration shows already on 1-2 days postoperatively one about 0.5 g / dl higher Hb value on. This is a sign of the known impact of the EPO to evaluate. In parallel, however, it comes to liver regeneration. Multiply here not only the hepatocytes but all cell types and above all the connective tissue structures that make up the architectural framework of the liver represent.

Experimental procedure:

28 Female pigs (weight 40.0-62.0 kg) were randomly divided into three groups. The partial removal of the left-sided Liver was performed with the technique of abdominal endoscopy.

Of the Cont Group (n = 16) was given no EPO. The group 2 (n = 6) was a combination of 10,000 units of EPO and a fibrin sealant (Quixil) are given locally to the liver resection surface. Group 3 was equally treated; however, the pigs received in addition to the local EPO treatment 10,000 units of EPO systemic on days 0, 3, 7 and 11.

liver samples were removed from the excised piece of liver on day 0, 24 hours after Resection from an area 1 cm below the resection surface and 14 days after resection below the resection surface and taken from the right lateral lobe.

For the proof of Ki-67 antigen, PCNA (proliferating cell nuclear antigen = PZNA Cell nuclear antigen proliferation) and apoptosis, was streptavidin-biotin Immunoperoxidase assay on formalin-fixed and in paraffin wax embedded liver tissue performed.

4. Optimized endoprosthesis or implants

implants can made of biodegradable but also of permanent materials consist. An example of this are metallic endoprostheses e.g. in the hip area. In connection to the production of the metallic raw form is a Mikrostrukturieurng by abrasion, etching or laser treatment achieved. This allows open porosities and Roughnesses in the range of 1 to 50-60 microns are generated.

These Structures then become in a solution made up of phase-pure beta tricalcium phosphate, giving a homogeneous surface coating is reached. After that, the structures will ideally become more advanced Sintering process to consolidate the beta-TCP structures.

Soluble salts / sugars can be incorporated in the mineral structures, or gas formation can be induced in order to achieve further interconnecting porosities. Subsequent to this manufacturing process, the structures are impregnated or coated with the growth factor or its derivatives, parts or analogues according to the invention. Deposits can be set accordingly on the surfaces by filling the cavities with EPO or using slow release substances. Alternatively, the implant with EPO and EPO can be taken out of the packaging immediately before implantation whose analogs are coated.

It This contributes to an improved fabric connection process the implant. The bone is connected macrovascularly and that implant accelerated and more sustainable integrated.

In similar Can shape Implants are prepared in the oral, maxillofacial and facial areas (Dental implants).

A Connection with a cellular Colonization in bioreactors with stem cells is possible.

biological Implants (vessels, heart valves, Skin) and membranes also be coated with EPO and or GH and or TPO.

5. Treatment of osteoporosis

EPO Coated tricalcium phosphate granules are detected by means of a needle puncture in deficit / rarefied vertebra in an autologous plasma solution brought in. There the granules are accelerated in endogenous bones rebuilt and transformed the degeneration process into an anabolic effect.

Of the Effect can be at risk of burglary vertebra be used. A combination with a colonization process advantageously with stem cells from the autologous bone marrow or the periosteum and blood and / or fatty tissue.

6th indication cartilage regeneration

chondrocytes represent strongly bradytrophic tissues. Through a regional trauma and abrasion cause inflammatory processes, that lead to arthitis can. By administration of EPO into the joint space or systemically and / or in Combination with cellular Transplants of chondrocytes or osteochondral cylinders promoted tissue regeneration and structural rebuilding. The combined systemic or subcutaneous administration of 10000 IU / day possible.

7. Indication skin diseases

Bad healing wounds are after preparation of the wound bed according to the invention with EPO or TPO (derivatives, analogs, parts) coated. This is preferred performed a mechanical debridement. To be in the blood clot 10 000 IE EPO. This process can be up to the cleaning of the wound ground.

The Structural growth begins after 2-3 days leading to accelerated training a granulation tissue.

8. Indication inflammatory bowel disease

at inflammatory phenomena of the intestinal tract with weight loss and anemia has been found that the anemia not a secondary phenomenon The reason for the poor food absorption is, but a concomitant phenomenon an original one Lack of EPO can be. The administration of EPO in 10 000 IU / day leads to a Improvement of intestinal repair / regeneration.

9th indication neuroregeneration

To Druchtrennungen of the spinal cord leads EPO to a structural growth of the neurons and re-sprouting the axons. supportive the administration of vitamin C.

EPO Can be regionally associated with fibrin glue / autologous plasma and / or endogenous Stem cells (bone marrow, blood CD 34, from fat cells, periosteum, umbilical cord) are given.

10. Parkinson's disease / example a chronic disease with already decayed inflammatory response

Autologous macrophages that have been stimulated by stimulation with degradable particles stereotactically integrated into degenerate areas. In parallel, EPO (10 000 IU) is injected into the area in conjunction with autologous stem cells (0.3 ml). Parallel to this, EPO is administered systemically over 2 weeks. This stimulation principle of macrophage inflammation induction can also be used in other chronic diseases where there is no acute trauma or acute inflammatory response.

Claims (32)

Use of hematopoietic growth factors, in particular erythropoietin (EPO) or thrombopoietin (TPO), or their derivatives, analogs or parts for the manufacture of a medicament to promote the structural regeneration of tissue. Use according to claim 2, characterized by the regeneration of traumatized tissue. Use according to claim 1 or 2, characterized by the use of the receptor-binding domain of growth factors or their derivatives or analogs. Use according to claim 1 or 2, characterized that a mimetic peptide is one of the growth factors, in particular EMP or DMP is used. Use according to one of Claims 1 to 4, characterized that the growth factors or their derivatives, analogs or parts across from the native growth factor additional Have glycosylation sites. Use according to one of claims 1 to 5, characterized that the growth factors or their derivatives, analogs or parts are conjugated with PEG. Use according to claim 1 or 2, countersigned by using the Novel Erythropoiesis Stimulating Protein (NESP). Use of an EPO inducing factor for production a drug for promotion structural tissue regeneration. Use according to one of claims 1 to 8, characterized that in addition one of the following factors is used: somatostatin, leukemia Inhibitory Factor (LIF), "Ciliary Neurotropic Factor "(CNTF)," Transforming Growth Factor beta "(TGF beta), prostaglandins, granulocyte-macrophage stimulating factor (GM-CSF), granulocyte-stimulating factor (G-CSF), growth hormone Releasing Hormones (GHRH), Thyrotropin-Releasing Homone (TRH), Gonadotropin-releasing Hormones (GnRH), Corticotropin Releasing Hormone (CRH), Dopamine, Antidiuretic hormone (ADH), oxytocin, prolactin, adrenocorticotropin, beta-Celltropin, Lutrotropin, vasopressin, nerve regeneration factors, preferably Nerve Growth Factor (NGF), vascular regeneration factors, Preferably, Vascular Endothelial Growth Factor (VEGF) or Plateled Derived Growth Factor (PDGF). Use according to one of claims 1 to 9, characterized that endothelial cells are present. Use according to one of claims 1 to 10, characterized that the regeneration of the tissue is directed locally. Use according to one of claims 1 to 10, characterized that the drug contains one or more of the factors after one of the claims 1 to 9 is administered topically. Use according to one of claims 1 to 10, characterized that the drug contains one or more of the factors after one of the claims 1 to 9 is systemically administered. Use according to one of claims 1 to 13, characterized that the growth process is supported by a support structure. Use according to claim 13, characterized that the support structure is treated with one of the factors according to one of claims 1 to 9. Use according to claim 14 or 15, characterized that as a carrier structure an implant, a graft or a carrier material for the growth of Cells is used. Use according to one of Claims 14 to 16, characterized that the support structure with Cells, preferably tissue-specific cells, progenitor cells, Bone marrow cells, peripheral blood, adipose tissue or fibrous tissue pre-populated or for in vivo colonization or inductive remodeling in vitro was prepared. Use according to claim 17, characterized that adult cells are progenitor cells, tissue-specific cells, preferably osteoblasts, fibroblasts, hepatocytes or smooth Muscle cells are used. Use according to one of claims 1 to 18, characterized that are during Encapsulated in the regeneration process forming cell aggregates and optionally frozen. Use according to one of claims 1 to 19 through the regeneration of nerve, muscle, epithelial or connective tissue as well as derived organs and structures. Use according to one of claims 1 to 20 for regeneration the liver, especially in cirrhosis, hepatitis, acute or chronic liver failure. Use according to one of claims 1 to 20 through the treatment for the regeneration of bones, cartilage, as well Tissues of endocrine organs, the heart muscle, heart valves, venous valves, Arterial valves, skin, vessels, aortas, Tendons, comea, tracea, nerves, miniscus, discus intervertebralis, Intestinal epithelium, ureters, urethra or bladder, as well as for treatment of degenerative diseases or to support tissue regeneration in chronic inflammation such as. in Crohn's disease, ulcerative colitis of diabetic ulcers, Gingiva or to stimulate neovascularization following a tissue injury. support structure containing at least one hematopoietic Growth factor or its derivatives, analogs or parts after one the claims 1 to 7 or an EPO inducing factor according to claim 8. support structure according to claim 23 additionally containing at least one of the following growth factors: somatostatin, Leukemia Inhibitory Factor (LIF), Ciliary Neurotropic Factor (CNTF), Transforming Growth Factor beta "(TGF beta), Prostaglandins, granulocyte macrophage stimulating factor (GM-CSF), Granulocyte-stimulating factor (G-CSF), growth hormone releasing hormone (GHRH), thyrotropin-releasing homone (TRH), gonadotropin-releasing Hormones (GnRH), Corticotropin Releasing Hormone (CRH), Dopamine, Antidiuretic hormone (ADH), oxytocin, prolactin, adrenocorticotropin, beta-Celltropin, Lutrotropin, Vasopressin, Nerve Regeneration Factors, preferably Nerve Growth Factor (NGF), vascular regeneration factors, preferably Vascular Endothelial Growth Factor (VEGF) or Plateled Derived Growth Factor (PDGF). support structure according to claim 23 or 24, characterized in that the support structure an implant, a graft, a carrier material for the growth of Cells, a stent, a patch, a catheter, skin, a hydrogel, a bone substitute material, an allogenic, autologous or xenogeneic, acellularized or non-acellularized tissue, a synthetic Tissue, a feeder layer or a flow is. support structure according to one of the claims 23 to 25, characterized in that the support structure with cells, preferably tissue-specific cells, progenitor cells, Bone marrow cells, peripheral blood, adipose tissue or fibrous tissue is pre-populated. support structure according to one of the claims 23 to 25, characterized in that the growth factors or their derivatives, analogs or parts according to any one of claims 1 to 9 are embedded in a biodegradable polymer layer. Process for producing a carrier structure for the Cell regeneration, characterized in that the support structure with at least one of the growth factors, their derivatives, analogues or parts according to any one of claims 1 to 9 is coated. A method according to claim 28, characterized by the activation of the support structure, preferably by plasma ionization or laser irradiation. A method according to any one of claims 28 and 29, characterized in that the support structure in vitro with cells, preferably tissue-specific cells, progenitor cells, bone marrow cells, peripheral Blood, fatty tissue or fibrous tissue is pre-colonized. Use according to one of claims 1 to 22, characterized that at least some of the process steps for promotion the tissue regeneration are carried out in whole or in part in vitro. Use according to claim 31, characterized that the growth process by stem cell delivery from the bone marrow, Blood, tissue, adipose tissue, umbilical cord tissue or blood.