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Bone Healing

Study Result Reference

Bone Healing

GHK-Cu was tested for healing of bone tissue

GHK-Cu markedly stimulated bone repair
Pickart L, US Patent 5,059,588   New and known copper peptide complexes for bone healing containing glycyl-histidyl-lysine and lysyl-histidyl-glycine.
Functions of bone forming cells. Effect of GHK-Cu on bone forming cells
GHK-Cu  increases the number of human marrow stromal cells and promotes the attachment of human osteoblastic cells.
Effects of the tripeptide  glycyl-L-histidyl-L-lysine copper complex on osteoblastic cell spreading, attachment and phenotype.  Godet and Marie (INSERM, Cell and Molecular Biology of Bone and Cartilage, Lariboisiere Hospital, Paris, France)  Cell Mol Biol (Noisy-le-grand) 1995 41(8):1081-91

Chick Bone forming cells 

Growth of bone chondrocytes

GHK-CU increased chondrocyte growth and their synthetic rate of bone collagen. Authors suggests GHK-Cu may be useful in the preparation of cartilage implants. 
Effect of the tripeptide glycyl-L-histidyl-L-lysine on the proliferation and synthetic activity of chick embryo chondrocytes. Pesakova, Novotna, and Adam (Institute of Rheumatology, Postgraduate Medical School, Czech Republic)  Biomaterials 1995 16(12):911-5

Bone Healing

The effect of GHK-Cu on the stimulation of new bone production in guinea pigs was studied.The authors prepared 7.5% and 12.5% collagen gels, supplemented with the tripeptide GHK-Cu, perfloxacine and hypersulphated glycosaminoglycan (HSGAG).

The gels were tested in guinea pigs for filling artificially created bone defects in diaphyses of femurs, and with cementless endoprostheses. Bone healing process was followed by means of RTG and NMR, and histologically. The slowest healing process was found in unfilled bone defects. Defects filled with Colladel without GHK-Cu healed more quickly, and the most accelerated healing was with the gels with GHK-Cu. The optimum gel, when used with cementless endoprostheses, produced vivid osteogenic activity at the interface of trabecular bone and metal stem. 
Morphological features of bone healing under the effect of collagen-graft-glycosaminoglycan  copolymer supplemented with the tripeptide Gly-His-Lys.  Pohunkova, Stehlik,  Vachal, Cech and Adam,  Institute of Rheumatology, Charles University, Prague, Czech Republic.  Biomaterials 1996, 17(16):1567-74 

 

Acta Chir Orthop Traumatol Cech. 1995;62(2):76-85. [The role of collagen implants containing the tripeptide gly-his-lys in bone healing process.]. [Article in Czech] Adam M1, Cech O, Pohunková H, Stehlík J, Klézl Z.

The authors prepared 12,5% collagen gel, that they supplemented by tripeptide gly-his-lys. peflacin and hypersulfated glycosaminoglycan (HSGAG). By means of I125 marking they followed up its absorption from small polyurethan sponges placed under the skin of rats. They found out, that absorption of gel without HSGAG is faster. No collagen antibodies (type I., II., III.) against the gel were formed neither in the experiment with rabbits, nor later on with minipigs when gel was placed into bone defects. Further on presence of microorganisms in the gel was looked for with negative results on various bacteriological nutrient media. The gel was used in a model experiment for filling artifically created bone defects in femurs of minipigs. Healing was followed up both histologically and with help of RTG and NMR and compared with healing of defects either unfilled, or filled with gel containing peflacin and HSGAG, but not tripeptide. The authors found out that slowest healing was in case of unfilled defects, on the other hand defects filled with collagen gel and GHK were healing substantially faster than defects filled with gel without GHK, i.e. healing needs approximately half the time less.

Acta Chir Orthop Traumatol Cech. 1995;62(6):336-42. [The effect of collagenous gel on endoprosthesis anchoring.]. [Article in Czech] Adam M1, Pohunková H, Cech O, Váchal J.

Collagen supplemented with tripeptide (gly-his-lys) was tested for implantation of cervicocapital endoprostheses of the hip in adult minipigs. In anaesthesia bone marrow cavity was filled up with the collagenous gel and then one of the two endoprosthesis types, produced by BMT Poldi Kladno was implanted. The experimental animals were killed eight, nine and ten weeks after operation. The femors were removed and roentgenologically and histologically examined. When bone marrow cavity was filled up with collagenous gel, anchoring of endoprosthesis with newly formed tissue was accelerated in comparison with those without gel. In the presence of fatty bone marrow residues healing process was disturbed. It cannot be excluded that this fact might have an effect on long term effective endoprosthesis anchoring.

Acta Chir Orthop Traumatol Cech. 1997;64(4):207-11. [Use of Bioimplants to Replace Cartilage Part II: Application of Implants in Animal Experiments.]. [Article in Czech] Adam M1, Pohunková H, Klézl Z, Pesáková V, Cech O.

The published paper pertains to a group of fullgrown minipigs where under anaesthesia with Narcamone after premedication with Rometar cartilage samples from the medial condyle of the femur in the area of the femoropatellar joint were taken. The collected cartilage samples served subsequently for the preparation of an autologous implant. After its preparation another operation was performed where by means of a bioptic trocar (diameter 3.5 mm) a cylindershaped defect was produced into which the implant was introduced. The animals were killed 8-12 weeks after the operation and part of the joint with the implant was subjected to further examination. The authors investigated three types of implants: 1. an implant based on three-dimensional cultivation of chondrocytes in a cultivation medium enriched with cartilaginous collagen and aggregan, 2. as sub 1 but the cultivation medium was enriched with the GHK tripeptide, 3. chondrocytes in the implant are added to gel prepared from cartilaginous collagens and aggregan. All three types of implants had a stimulating effect on the new formation of hyaline cartilage. When the tripeptide GHK is used, the newly formed tissue contains more cells. Simplest is the preparation of the third implant. Also the peroperative procedure is simplest. Key words: chondrocytes, cartilaginous collagens, cartilaginous implant, tripeptide gly-his-lys, aggregan, minipigs.

GHK-Cu stimulates bone healing in animals and the functions of bone repair cells in culture. The development of GHK-Cu for clinical use is being conducted under the direction of Prof. Milan Adam (University of Prague, Photograph - on the left).
 

This paper uses a mixture of 3 peptides, at relatively low dosages, to heal bone fractures.

The people at the University of Kursk have found an improved method in the use of GHK. They used Gly-His-Lys (0.5 μg/kg), dalargin (1.2 μg/kg) (an opioid-like synthetic drug) and and biological peptide thymogen (0.5 μg/kg) (dipeptide,L-glutamyl-L-tryptophan) to heal bones. The total peptide dosage is about 2 micrograms per KG or about 140 micrograms in a human. 

Synergetic antioxidant and reparative action of thymogen, dalargin and peptide Gly-His-Lys in tubular bone fractures. Cherdakov V.Yu., Smakhtin M.Yu., Dubrovin G.M., Dudka V.T., Bobyntsev I.I. , Experimental Biology and Medicine, No.4. 2010 (In Russian)

Biochemical Department, Traumatology, Orthopedics & Military Surgery Department, Pathanatomy Department, Pathophysiology Department of the Kursk State Medical University, Kursk

E-mail: cherdakov@rambler.ru

It has been established, that in case of experimental rat bone fractures the intraperitoneally injections of peptides in equimolar doses – Gly-His-Lys (0.5 μg/kg), dalargin (1.2 μg/kg) and thymogen (0.5 μg/kg) during 10 days result in their antioxidant actions testified by the decrease of malonic dialdehyde concentration and increase of catalase activity in blood. The effect of thymogen on the level of malonic dialdehyde was more expressed in comparison with Gly-His-Lys action. The peptides had synergetic and reparative action because in case of twin combined injections of these substances the essential amplification of antioxidant (catalase activity) and reparative effects was observed in comparison with individual injections of peptides. The more expressed reparative changes were found in the group of rats with peptide combined injection Gly-His-Lys + dalargin testifyed by histological and X-ray investigations. In animals of this group the line of bone fracture was not observed after 10 days practically. The synergetic action of peptides Gly-His-Lys, thymogen and dalargin may be used for stimulation of reparative osteogenesis.


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