Exogenous Application Of Growth Factors Research Articles

Osteogenesis and Chondrogenesis of Primary Rabbit Periosteal Cells under Non-uniform 2-Axial Tensile Strain

Periosteal cells are the major cell sources of skeletal progenitors for fracture callus. In order to promote bone repair and cartilage formation, besides the application of exogenous growth factors, physical stimulation is an alternative approach to guide cell differentiation. Investigation of appropriate conditions is essential for forming bone and cartilage. In this work, a membrane-type micro-system was developed to provide cell culture environment and cell stretching stimulation during culture course. Circular and oval culture wells were designed to respectively generate uniform and non-uniform 2-axial tensile strain for stretching primary rabbit periosteal cells. Cell orientation and differentiation were studied after cycling stretching for 2 days. The cells aligned to the stretching axis with high tensile strain in the oval culture wells; while the cells expressed random orientation in the circular culture wells. Different responses were significantly shown when the cells were respectively stimulated by uniform and non-uniform 2-axial tensile strains. On the other hand, osteogenic differentiation was shown when the cells were under either uniform or non-uniform 2-axial tensile strain. However, only non-uniform 2-axial tensile strain could induce mature osteoblasts. In addition, the result revealed chondrocytes could be differentiated only under a large and nearly single dimensional tensile strain. In summary, differentiation of the periosteal cells is highly influenced by 2-dimensional distribution of the tensile strain. This work provides some in-sights of the control of axial tensile strain for periosteal cell differentiation.

Syndecan -4 and Alkaline Phosphatase Enhancement by Local Application of Exogenous Growth Factors on Traumatic Pulp of Osteoporotic Rats: Immuno-Histochemical Analysis

Syndecan -4 and Alkaline Phosphatase Enhancement by Local Application of Exogenous Growth Factors on Traumatic Pulp of Osteoporotic Rats: Immuno-Histochemical Analysis

Role and influence of growth factors on early osseointegration in animal jaw bone: A meta-analysis.

Aim:Growth factors (GFs) are polypeptides, which are intricately involved in the regulation of bone formation, preservation, and regeneration through gene expression. However, the role of these bioactive agents in osseointegration of dental implants has not been substantially proven. The objective of this systematic review (SR) and meta-analysis was to explore the effect of GFs on early osseointegration of dental implants in animal jaws. An attempt to decipher an adjunctive role of GFs in modulating predictable bone growth in peri-implant areas was done.Materials and Methods:An electronic and manual search of different databases was performed. Only randomized controlled trials (RCTs) were included and reviewed. The risk of bias (ROB) of the selected studies was assessed using the SR Centre for Laboratory Animal Experimentation (Cochrane) tool. A meta-analysis was also performed to evaluate the different study characteristics quantitatively.Statistical Analysis used:The total Weighted mean difference was evaluated using the Rev-Manv5.3 algorithm. Chi-square test and I2 test were done to assess the heterogeneity between the studies.Results:Seven RCTs were included in the study. These were associated with a high ROB. The total weighted mean difference (WMD) of the percentage of bone–implant contact was 3.25% (95% confidence interval [CI] = 1.49%–6.03%; P = 0.001; I2 = 91%) between groups with and without exogenous application of GFs. The total WMD of the percentage of newly formed bone area was 4.48% (95% CI = 2.31%–5.90%; P < 0.00001, I2 = 84%). A high level of heterogeneity (P < 0.001 for Chi-square test; I2>50 %) among comparable studies was observed.Conclusion:The ancillary application of external GFs exhibited evidence of early osseointegration, resulting in more predictable and faster results. However, a careful discernment of conclusions drawn from this SR is a must before conducting any human trials.

Advanced Growth Factor Delivery Systems in Wound Management and Skin Regeneration.

Growth factors are endogenous signaling molecules that regulate cellular responses required for wound healing processes such as migration, proliferation, and differentiation. However, exogenous application of growth factors has limited effectiveness in clinical settings due to their low in vivo stability, restricted absorption through skin around wound lesions, elimination by exudation prior to reaching the wound area, and other unwanted side effects. Sophisticated systems to control the spatio-temporal delivery of growth factors are required for the effective and safe use of growth factors as regenerative treatments in clinical practice, such as biomaterial-based drug delivery systems (DDSs). The current review describes the roles of growth factors in wound healing, their clinical applications for the treatment of chronic wounds, and advances in growth factor-loaded DDSs for enhanced wound healing, focusing on micro- and nano-particulate systems, scaffolds, hydrogels, and other miscellaneous systems.

Pressure Ulcer Research: Current and Future Perspectives

This multi-author volume brings together 30 contributors under an international editorship. The four editors include three biomedical engineers (one from Queen Mary's College, University of London and two from the Eindhoven University of Technology) and the medical director of l'Arche Rehabilitation Centre in France, who is the only clinical author. The final chapter on tissue repair strategies, which is of most surgical interest, focuses on ‘biochemical stimulation of the wound bed to improve wound healing. Important new therapeutics in this category are reviewed such as: (i) exogenous application of growth factors; (ii) tissue-engineered skin grafts; and (iii) gene therapy.’ The two authors from the Eindhoven University of Technology are (according to an internet search) not medically qualified, so understandably their viewpoints do not set out the full range of current reconstructive surgical options or ‘future perspectives’ in clinical surgery. The publishers have adopted the convention of excluding any reference to the contributors' professional qualifications. With this handicap, the clinical reader cannot effectively assess treatment strategies mentioned by the authors as merely being references from the literature or from the author's own authoritative experience. Nevertheless, this is a useful and unique contribution to a neglected area. Risk assessment, basic science and human and animal research are fully reviewed. Epidemiological studies from The Netherlands on the costs of pressure ulcer care and the medicolegal review from the US add significant practical insights. The chapter on the role of oxidative stress in the development and persistence of pressure ulcers is a particularly helpful summary of ischaemia-reperfusion injury from two Oxford biochemists who support their chapter with a comprehensive reference list. The editors have generally avoided repetition (a significant achievement in a multi-author text) though there is a colour illustration of a section through human skin that is a reiteration of a black and white graphic. This slim volume succeeds in its aim of providing ‘a clearer understanding of the mechanisms associated with the aetiology of pressure ulcers and to identify the risk levels of individuals’. Surgeons with a pressure sore and wound-healing interest will find a useful reference even if it does not find wider surgical acceptance. I would certainly want to have this on my medical bookshelf.

Local Regeneration of Dentin-Pulp Complex Using Controlled Release of FGF-2 and Naturally Derived Sponge-Like Scaffolds

Restorative and endodontic procedures have been recently developed in an attempt to preserve the vitality of dental pulp after exposure to external stimuli, such as caries infection or traumatic injury. When damage to dental pulp is reversible, pulp wound healing can proceed, whereas irreversible damage induces pathological changes in dental pulp, eventually requiring its removal. Nonvital teeth lose their defensive abilities and become severely damaged, resulting in extraction. Development of regeneration therapy for the dentin-pulp complex is important to overcome limitations with presently available therapies. Three strategies to regenerate the dentin-pulp complex have been proposed; regeneration of the entire tooth, local regeneration of the dentin-pulp complex from amputated dental pulp, and regeneration of dental pulp from apical dental pulp or periapical tissues. In this paper, we focus on the local regeneration of the dentin-pulp complex by application of exogenous growth factors and scaffolds to amputated dental pulp.

Regeneration Approaches for Dental Pulp and Periapical Tissues with Growth Factors, Biomaterials, and Laser Irradiation

In current dental practice, restorative and endodontic procedures have been developed in an attempt to preserve the vitality of dental pulp after exposure to external stimuli such as caries infection. When damage to dental pulp is reversible, pulp wound healing can proceed, whereas irreversible damage induces pathological changes in dental pulp, eventually requiring its removal. Furthermore, dentists sometimes extract non-vital teeth because of severe caries progression, critical size of periapical lesion, and tooth fracture. To overcome the limitations of presently available therapies, it is important to develop regeneration therapy for dental pulp and periapical tissues. In this review, we focus on the regeneration of dental pulp and periapical tissues by application of exogenous growth factors and scaffolds, as well as low-intensity laser irradiation as an auxiliary therapy for regeneration therapy.

Schwann Cells as a Therapeutic Target for Peripheral Neuropathies

Schwann cells, the myelin forming cells in the peripheral nervous system, play a key role in the pathology of various inflammatory, metabolic and hereditary polyneuropathies. Advances in identifying growth factors and signaling molecules that are expressed by Schwann cells have paved the way for development of new treatment strategies that are aimed to improve the protective and regenerative properties of Schwann cells in peripheral nerve disorders. These include the exogenous application of growth factors and neurohormones which have been advanced into clinical trials in humans, and transplantation paradigms that have been moved into late stage preclinical models. In this review we will discuss the latest developments in these therapeutic approaches with special regard to peripheral nerve disorders, in which progress in basic research has already been translated into clinical trials, including HIV-associated distal sensory polyneuropathy and diabetic neuropathy.

The impact of hyperbaric oxygen therapy on serological values of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF)

BackgroundHyperbaric oxygen (HBO) therapy is an effective adjunct treatment for ischemic disorders such as chronic infection or chronic wounds. It combines hyperoxic effects with the stimulating potential of post-therapeutic reactive hypoxia. As its crucial effects, stimulation of fibroblast growth, induction of collagen synthesis and the initiation of angiogenesis are discussed. Angiogenesis is a multistage process resulting in the growth of blood vessels. It includes degradation of extracellular matrix, proliferation and migration of different cell populations and finally formation of new vessel structures. This complex chain of procedures is orchestrated by different cytokines and growth factors. Crucial mediators of angiogenesis are basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF); their in-vivo function is still not fully understood.MethodsForty-three patients suffering from sudden sensorineural hearing loss or tinnitus were treated with HBO. The therapy included 10 sessions of 90 minutes each, one session a day. Serological levels of bFGF and VEGF were assessed by enzyme-linked immunosorbent assays performed according to the manufacturer's instructions on day 1, 2, 5 and 10 of HBO therapy and were compared to mean values of the control group, related to the patient's age and sex, and their development observed over the ten days of HBO.ResultsThere was no sex- or age dependency of bFGF observed in the present study, whereas under HBO our results showed a significant mitigation of the bFGF concentration. In the present data, there was no connection between the VEGF concentration and the patients' ages. Women showed significantly higher levels of VEGF. There was no significant change of VEGF concentration or the VEGF/bFGF ratio during HBO. All scored results varied within the range of standard values as described in the current literature.ConclusionsA significant effect of HBO on serum concentrations of bFGF and VEGF was not verified in the present study. Additional application of exogenous growth factors in conjunction with HBO was not obviously linked by a coherent cause-and-effect chain as far as wound healing is concerned.

Keloids: Fundamental principles and prospects (Review)

Wound healing is a very complex process of interactions between different cells, growth factors, blood elements and extracellular matrix. Keloids represent one of the possible complications in the fundamental process of cutaneous wound repair. Despite all efforts, keloids remain a therapeutic challenge since no treatment is as yet considered 100% effective. Growth factors, discovered in the late 1970s, have been shown to influence dermal regeneration. However, the exogenous application of growth factors to chronic wounds has not proven to be effective in healing them. Additionally, genetic analysis has not revealed any single gene that might cause keloids; as such, classic gene therapy is not a feasible option for the treatment of keloids. A new approach is so-called somatic gene therapy. This review provides an overview of the fundamentals of wound healing and of keloids, and presents new possibilities that may improve cutaneous wound repair.

Growth factors improve neurogenesis and outcome after focal cerebral ischemia

Stem cells have been proposed as a new form of cell-based therapy in a variety of disorders, including acute and degenerative brain diseases. Endogenous neural stem cells (eNSC) reside in the subventricular zone and in the subgranular zone of the hippocampus. eNSC are capable of self-renewal and differentiation into functional glia and neurons. Unfortunately, spontaneous brain regeneration is inefficient for clinically significant improvement following brain injury. However, eNSC responses may be augmented considerably by perturbing the pathways governing cell proliferation, migration and differentiation by application of exogenous growth factors. Importantly, current evidence suggests that such perturbations may lead to better functional outcome after stroke. This article summarizes the progress made in this field.

Role of connective tissue growth factor and its interaction with basic fibroblast growth factor and macrophage chemoattractant protein‐1 in skin fibrosis

Activation of the immune system and abnormal growth of skin fibroblasts cause systemic sclerosis. Growth factors have various biological activities, including mediation of immune reactions. The growth factor family includes basic fibroblast growth factor (bFGF), transforming growth factor-beta (TGF-beta), and connective tissue growth factor (CTGF). CTGF, an important downstream mediator of TGF-beta in fibrosis, has been suggested to play a specific role in fibrotic disorders. We have directed our attention to the role of CTGF in sustaining skin fibrosis. To better understand its effects in vivo, we established an animal model of skin fibrosis induced by exogenous application of growth factors. In this model, bFGF transiently induced subcutaneous fibrosis. Simultaneous injection of bFGF and CTGF increased skin fibrosis compared with a single injection of bFGF. Serial injections of bFGF for 3 days followed by CTGF for 4 days, or of CTGF followed by bFGF, did not cause skin fibrosis but simultaneous injections increased macrophage chemoattractant protein-1 (MCP-1) mRNA expression levels. To further define the mechanisms of skin fibrosis in vivo, bFGF and CTGF were injected simultaneously into MCP-1 knockout mice, resulting in decreased collagen levels in granulation tissues on day 8. The number of inflammatory cells, such as mast cells, macrophages and lymphocytes, was significantly decreased in MCP-1 knockout mice compared with wild-type mice. These results suggest that bFGF induces collagen production by stimulating skin fibroblasts and CTGF cooperates with bFGF. Our results indicate that the induction of MCP-1 is necessary for infiltration of inflammatory cells.

Neutralizing monoclonal antibody to human connective tissue growth factor ameliorates transforming growth factor‐β‐induced mouse fibrosis

Skin fibrotic disorders such as systemic sclerosis (SSc) are characterized by an excessive accumulation of extracellular matrix (ECM) and are understood to develop under the influence of fibrogenic growth factors. To better understand the detailed mechanisms of persistent fibrosis in SSc, we have previously established an animal model of skin fibrosis induced by exogenous application of growth factors. In this model, transforming growth factor-beta (TGF-beta) transiently induced subcutaneous fibrosis and serial injections of connective tissue growth factor (CTGF) after TGF-beta caused persistent fibrosis. These results suggest that CTGF plays an important role in the development of persistent skin fibrosis and that CTGF may be a potential and specific therapeutic target in skin fibrosis. Therefore, the aim of the current study is to develop a neutralizing monoclonal antibody against human CTGF. We also investigated the neutralizing effect of the antibodies in our animal model. Firstly, by using the DNA immunization method, we developed a panel of anti-CTGF antibodies recognizing the native conformation of human CTGF. Next, to examine the anti-fibrosing effects of these antibodies, newborn B6 mice received subcutaneous injections of TGF-beta for 3 days with either anti-CTGF neutralizing antibodies or control purified immunoglobulin. Anti-CTGF antibodies significantly reduced skin fibrosis and collagen contents compared with the control group. These results suggest that our anti-CTGF antibodies are capable of blocking the development of skin fibrosis at least partially and these anti-CTGF neutralizing antibodies may be useful as the feasible strategy to treat skin fibrotic diseases as SSc.

Collective Review: Bioactive Implants Coated with Poly(D,L-lactide) and Growth Factors IGF-I, TGF-β1, or BMP-2 for Stimulation of Fracture Healing

Demographic data reveal that due to the increasing aging of the population, complications with the musculoskeletal system will increase in the next years. One major problem in orthopedic and trauma surgery are the delayed healing or non-unions of long bone fractures. The exogenous application of growth factors can stimulate the bone healing to reduce these complications. Beside the choice of the optimal growth factor the application system is important. Therefore, we developed a new bioactive coating method for implants, which is based on a biodegradable poly(D,L-lactide) (coating thickness: 10 mum). This coating allows the incorporation of growth factors and the controlled release of these factors during the healing process without the need for further devices. The effect of different growth factors (IGF-I, TGF-beta1, and BMP-2) locally released from coated intramedullary implants on fracture healing was investigated with biomechanical and histological analysis in rats. All investigated growth factors stimulated the fracture healing as assessed with biomechanical tests and histological analysis. The local application of combined IGF-I and TGF-beta1 had the most stimulating effect on fracture healing, followed by the effect of BMP-2, IGF-I, and TGF-beta1 alone. Bioactive coating of biomechanical well-established implants can on the one hand stabilize the fracture and on the other hand stimulate healing processes to increase healing and to reduce the rate of complications.

Wound Healing Enhancement: Electroporation to Address a Classic Problem of Military Medicine

The major goal of wound healing biology is to determine how a wound can be induced to repair damaged tissue faster and more efficiently. Enhancement of dermal and epidermal regeneration is an extremely important goal for the treatment of many different types of wounds. Exogenous application of growth factors to the wound site has been shown to have potential to improve wound healing. Frequent applications of large amounts of growth factor have been required. This is because proteases in the wound quickly destroy peptide growth factor. Gene therapy has the potential to produce growth factors deep within the wound, where they can be effective as well as able to constantly replenish growth factor that is destroyed by peptidases. We have shown that application of plasmid DNA expression vectors directly into the wound is an inefficient modality. Electroporation, the application of an electrical field across cells to permeabilize the cell membrane has led us to explore the possibility of utilizing the technique to enhance transfection efficiency. We have identified electroporation parameters that improve the efficiency of DNA transfection in cutaneous wounds, and we have shown that electroporation itself does not impair wound healing. We are now on the threshold of exploring whether electroporation-assisted transfection with DNA plasmid expression vectors for growth factors will be an effective modality for enhancing cutaneous wound healing.

Engineered growth factors and cutaneous wound healing: Success and possible questions in the past 10 years

In the past 10 years, many engineered growth factors, including recombinant human epidermal growth factor, basic fibroblast growth factor, and platelet-derived growth factor, have been produced and used in the clinic. After screening the results from different centers, some results are found to be encouraging, while others are discouraging. Although the interpretation of these results may depend on your perspective, it may also depend on different criteria, different wounds, and even different aims. In this article, successful experiences and failures concerning the use of growth factors and cutaneous wound healing are summarized. Based on this information and our clinical experience, we address people's concerns such as whether growth factors have altered clinical practice thus far and whether growth factor treatments have solved all problems involved in wound healing. Is there a need for exogenous application of growth factors in acute or chronic wounds, and if so, is it safe to use growth factors to promote wound healing? Last, can we achieve perfect wound healing in those wounds treated with growth factors?

BFGF and EGF modulate trauma-induced proliferation and neurogenesis in juvenile organotypic hippocampal slice cultures

Since postnatal and adult mammalian brains have been shown to retain an ability to generate neurons from endogenous stem cells throughout life, these cells could play a central role in regeneration after neuronal loss. Therefore, we studied cell proliferation, glio- and neurogenesis respectively after brain injury in organotypic hippocampal slice cultures using a focal trauma by transecting Schaffer collaterals in the cornu ammonis (CA) 2 region mechanically. After determination of cell death using propidium iodide, neuroregenerative processes were quantitatively analyzed by various immunohistochemical techniques at different time points post injury. As this endogenous insult-induced neurogenesis is rather inefficient, we investigated if it can be enhanced by application of exogenous growth factors. Exogenous basic fibroblast growth factor (bFGF) enhanced neurogenesis significantly in the dentate gyrus (DG) region. A neutralizing antibody against endogenous bFGF revealed a significant decrease of basal and trauma-induced proliferation. Reverse transcription polymerase chain reaction (RT-PCR) studies exhibited a downregulation of FGF messenger ribonucleic acid (mRNA) transcription after the antibody treatment. In contrast, epidermal growth factor (EGF) increased proliferation, but not neurogenesis. A combination of bFGF and EGF displayed an EGF-like effect on proliferation and no effect on neurogenesis. These results demonstrate, that in our model bFGF but not EGF sustains neurogenesis, whereas together the two growth factors permit an increased proliferation but not neurogenesis in organic hippocampal slice cultures.

Growth factors and skin repair and regeneration

Growth factors and skin repair and regeneration

Connective tissue growth factor causes persistent proα2(I) collagen gene expression induced by transforming growth factor‐β in a mouse fibrosis model

Skin fibrotic disorders such as systemic sclerosis (SSc) are characterized by an excessive production of extracellular matrix (ECM) and understood to develop under the influence of certain growth factors. Connective tissue growth factor (CTGF) is a cysteine-rich mitogenic peptide that is implicated in various fibrotic disorders and induced in fibroblasts after activation with transforming growth factor-beta (TGF-beta). To better understand the mechanisms of persistent fibrosis seen in SSc, we previously established an animal model of skin fibrosis induced by exogenous application of growth factors. In this model, TGF-beta transiently induced subcutaneous fibrosis and serial injections of CTGF after TGF-beta caused persistent fibrosis. To further define the mechanisms of skin fibrosis induced by TGF-beta and CTGF in vivo, we investigated in this study, the effects of growth factors on the promoter activity of the proalpha2 (I) collagen (COL1A2) gene in skin fibrosis. For this purpose, we utilized transgenic reporter mice harboring the -17 kb promoter sequence of the mouse COL1A2 linked to either a firefly luciferase gene or a bacterial beta-galactosidase gene. Serial injections of CTGF after TGF-beta resulted in a sustained elevation of COL1A2 mRNA expression and promoter activity compared with consecutive injection of TGF-beta alone on day 8. We also demonstrated that the number of fibroblasts with activated COL1A2 transcription was increased by serial injections of CTGF after TGF-beta in comparison with the injection of TGF-beta alone. Furthermore, the serial injections recruited mast cells and macrophages. The number of mast cells reached a maximum on day 4 and remained relatively high up to day 8. In contrast to the kinetics of mast cells, the number of macrophages was increased on day 4 and continued to rise during the subsequent consecutive CTGF injections until day 8. These results suggested that CTGF maintains TGF-beta-induced skin fibrosis by sustaining COL1A2 promoter activation and increasing the number of activated fibroblasts. The infiltrated mast cells and macrophages may also contribute to the maintenance of fibrosis.

Short term in vivo biocompatibility testing of biodegradable poly( D, L-lactide)—growth factor coating for orthopaedic implants

Fracture healing can be stimulated by exogenous application of growth factors. Using porcine and rat models the efficacy of locally delivered IGF-I and TGF- β1 from an implant coating has been demonstrated. A thin and biomechanical stable biodegradable poly( D, L-lactide) was used to coat implants and serve as a drug carrier. Due to reports of possible foreign body reactions caused by polymer materials in orthopedic surgery, this study investigated the biocompatibility of the polylactide implant coating and the locally released growth factors during the time course of rat tibial fracture healing (days 5, 10, 15, and 28 after fracture). Monocytes/macrophages and osteoclast were detected using an monoclonal antibody against ED1 (comparable to CD68 in mice and human). The antibody ED1 stains monocytes, macrophages and osteoclast in the bone marrow and in the newly formed fracture callus. A moderate density of the monocytes/macrophages was seen in the proximal part of the medullary canal, but almost no cells were detectable in the region distal to the fracture. The amount of stained cells increased during the observation time with a maximum at days 10 and 15 followed by a decrease at day 28. No differences were detectable between the investigated groups from day 5 to 15 post fracture indicating, that the used poly( D, L-lactide) or the incorporated growth factors do not evoke an elevated immunological response compared to the uncoated titanium implant at the investigated time points. A significantly higher amount of ED1 positive cells was measured 28 days after fracture in the control group compared to the groups with the coated implants. In conclusion, no indication of a foreign body reaction due to the use of the polylactide or the growth factors was found indicating a good short-term biocompatibility of this bioactive coating.