In vivo incisional wound healing augmented by platelet-derived growth factor and recombinant c-sis gene homodimeric proteins.

Human platelet-derived growth factor (hPDGF) is likely to be important in stimulating tissue repair, based upon its in vivo chemotactic and stimulatory activities for inflammatory cells and fibroblasts and upon the presence of PDGF and related proteins in platelets, macrophages, and activated fibroblasts, cell types that make up the milieu of the healing wound. Recombinant human c-sis (rPDGF-B), homodimers of the B chain of PDGF, were compared with hPDGF in vitro. rPDGF-B was immunologically similar to hPDGF and, at identical concentrations, similar to hPDGF in stimulating fibroblast mitogenesis and chemotaxis of polymorphonuclear leukocytes, monocytes, and fibroblasts. Purified hPDGF and rPDGF-B were also tested in vivo for potency in a model of tissue repair using a linear incision wound through rat dermis. A single application of hPDGF or rPDGF-B (2-20 micrograms/wound) in a slow release vehicle at the time of wounding resulted in a dose-dependent, statistically highly significant increase of breaking strength of treated wounds. Wound healing in animals treated with rPDGF-B was 170% stronger and accelerated by 2 d during the first week over control wounds and by 4-6 d over the next 2 wk. Histologic evaluation of growth factor-treated wounds correlated the in vitro chemotactic activity and the accelerated healing of wounds with a striking inflammatory cell infiltrate early after wounding, markedly increased formation of granulation tissue by 4-d, and increased fibrosis by 14 d in comparison to control wounds. The results thus demonstrate that rPDGF-B is fully active in in vitro tests of mitogenesis and chemotaxis and, for the first time, demonstrate directly that PDGF significantly advances wound healing in incisional wounds of experimental animals.

Platelet-derived growth factor and transforming growth factor-beta enhance tissue repair activities by unique mechanisms.

Platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta) markedly potentiate tissue repair in vivo. In the present experiments, both in vitro and in vivo responses to PDGF and TGF-beta were tested to identify mechanisms whereby these growth factors might each enhance the wound-healing response. Recombinant human PDGF B-chain homodimers (PDGF-BB) and TGF-beta 1 had identical dose-response curves in chemotactic assays with monocytes and fibroblasts as the natural proteins from platelets. Single applications of PDGF-BB (2 micrograms, 80 pmol) and TGF-beta 1 (20 micrograms, 600 pmol) were next applied to linear incisions in rats and each enhanced the strength required to disrupt the wounds at 5 d up to 212% of paired control wounds. Histological analysis of treated wounds demonstrated an in vivo chemotactic response of macrophages and fibroblasts to both PDGF-BB and to TGF-beta 1 but the response to TGF-beta 1 was significantly less than that observed with PDGF-BB. Marked increases of procollagen type I were observed by immunohistochemical staining in fibroblasts in treated wounds during the first week. The augmented breaking strength of TGF-beta 1 was not observed 2 and 3 wk after wounding. However, the positive influence of PDGF-BB on wound breaking strength persisted through the 7 wk of testing. Furthermore, PDGF-BB-treated wounds had persistently increased numbers of fibroblasts and granulation tissue through day 21, whereas the enhanced cellular influx in TGF-beta 1-treated wounds was not detectable beyond day 7. Wound macrophages and fibroblasts from PDGF-BB-treated wounds contained sharply increased levels of immunohistochemically detectable intracellular TGF-beta. Furthermore, PDGF-BB in vitro induced a marked, time-dependent stimulation of TGF-beta mRNA levels in cultured normal rat kidney fibroblasts. The results suggest that TGF-beta transiently attracts fibroblasts into the wound and may stimulate collagen synthesis directly. In contrast, PDGF is a more potent chemoattractant for wound macrophages and fibroblasts and may stimulate these cells to express endogenous growth factors, including TGF-beta, which, in turn, directly stimulate new collagen synthesis and sustained enhancement of wound healing over a more prolonged period of time.

Accelerated healing of incisional wounds in rats induced by transforming growth factor-beta.

The role of polypeptide growth factors in the processes of inflammation and repair was investigated by analyzing the influence of transforming growth factor-beta (TGF-beta), applied directly to linear incisions made through rat dorsal skin. A dose-dependent, direct stimulatory effect of a single application of TGF-beta on the breaking strength of healing incisional wounds was demonstrated. An increase in maximum wound strength of 220 percent of control was observed at 5 days; the healing rate was accelerated by approximately 3 days for at least 14 days after production of the wound and application of TGF-beta. These increases in wound strength were accompanied by an increased influx of mononuclear cells and fibroblasts and by marked increases in collagen deposition at the site of application of TGF-beta. TGF-beta is thus a potent pharmacologic agent that can accelerate wound healing in rats.

Growth factor-induced acceleration of tissue repair through direct and inductive activities in a rabbit dermal ulcer model.

The roles of polypeptide growth factors in promoting wound healing and in directing the specificity and sequence of responses of different tissues in wounds are little understood. We investigated the influence of four growth factors on the rates of healing of a novel full thickness dermal ulcer placed on an avascular base in the rabbit ear. The wound model precludes significant wound contraction and requires new granulation tissue and epithelial cells for healing to originate centripetally. 5 micrograms (7-31 pmol/mm2) of platelet-derived growth factor-B chain (PDGF-BB), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF) applied locally at the time of wounding resulted in a twofold increase in complete reepithelialization of treated wounds (PDGF-BB, P = 0.02 chi square analysis; bFGF, P = 0.04; EGF, P = 0.05); transforming growth factor (TGF)-beta 1 significantly inhibited reepithelialization (P = 0.05). Both PDGF-BB and TGF-beta 1 uniquely increased the depth and area of new granulation tissue (P less than 0.005), the influx of fibroblasts, and the deposition of new matrix into wounds. Explants from 7-d old PDGF-BB-treated wounds remained metabolically far more active than controls, incorporating 473% more [3H]thymidine into DNA (P = 0.05) and significantly more [3H]leucine and [3H]proline into collagenase-sensitive protein (P = 0.04). The results establish that polypeptide growth factors have significant and selective positive influences on healing of full thickness ulcers in the rabbit.

Interaction between the insulin-like growth factor family and the integrin receptor family in tissue repair processes. Evidence in a rabbit ear dermal ulcer model.

We have determined previously that IGF-I is dependent on the presence of IGF binding protein-1 (IGFBP-1) to act as a wound healing agent. We sought to determine the mechanism whereby IGFBP-1 is able to enhance IGF-I bioactivity. As IGFBP-1 binds both the alpha5beta1 integrin as well as IGF-I in vitro, we asked which of the following interactions were important: (a) the ability of IGFBP-1 to interact with an integrin receptor, and/or (b) the binding of IGF-I by IGFBP-1. We used an IGF-1 analogue (des(1-3)IGF-I) with a > 100-fold reduction in affinity for IGFBP-1 as well as an IGFBP-1 mutant (WGD-IGFBP-1) which does not associate with the alpha5beta1 integrin to selectively abrogate each of these interactions. We also tested the ability of IGFBP-2, a related binding protein which has an arginine-glycine-aspartate sequence but does not associate with integrin family members, to enhance IGF-I bioactivity. Full-thickness dermal wounds were created on rabbit ears; various combinations of native IGF-I, native IGFBP-1, native IGFBP-2, and their respective analogues/mutants were applied to each wound. Wounds were harvested 7 d later for analysis. Only native IGF-I in combination with native IGFBP-1 was effective as a wound healing agent, enhancing reepithelialization and granulation tissue deposition by 64+/-5 and 83+/-12% over controls (P = 0.008 and 0.016, respectively). The same doses of IGF-I/WGD-IGFBP-1, des(1-3)IGF-I/IGFBP-1, and IGF-I/IGFBP-2 were ineffective. We propose that IGF-I physically interacts with IGFBP-1 and that IGFBP-1 also binds to an integrin receptor, most likely the alpha5beta1 integrin. This interaction is unique to IGFBP-1 as the closely related IGFBP-2 had no effect, a finding consistent with its inability to bind to integrin receptors. Our results suggest that activation of both the IGF-I receptor and the alpha5beta1 integrin is required for IGF-I to stimulate wound healing.

Hypoxia increases human keratinocyte motility on connective tissue.

Re-epithelialization of skin wounds depends upon the migration of keratinocytes from the cut margins of the wound and is enhanced when human keratinocytes are covered with occlusive dressings that induce hypoxia. In this study, two independent migration assays were used to compare cellular motility on connective tissue components under normoxic or hypoxic conditions. Human keratinocytes apposed to collagens or fibronectin exhibited increased motility when subjected to hypoxic (0.2 or 2% oxygen) conditions compared with normoxic (9 or 20% oxygen) conditions. When compared with normoxic cells, hypoxic keratinocytes exhibited increased expression and redistribution of the lamellipodia-associated proteins (ezrin, radixin, and moesin). Furthermore, hypoxic keratinocytes demonstrated decreased secretion of laminin-5, a laminin isoform known to inhibit keratinocyte motility. Hypoxia did not alter the number of integrin receptors on the cell surface, but did induce enhanced secretion of the 92-kD type IV collagenase. These data demonstrate that hypoxia promotes human keratinocyte motility on connective tissue. Hypoxia-driven motility is associated with increased expression of lamellipodia proteins, increased expression of collagenase and decreased expression of laminin-5, the locomotion brake for keratinocytes.

Differential activation of migration by hypoxia in keratinocytes isolated from donors of increasing age: implication for chronic wounds in the elderly.

Chronic wound healing conditions are often observed in elderly patients with poor tissue oxygenation. Impaired re-epithelialization is a hallmark of these wounds, which is seen in both clinical studies and in our animal models of impaired healing. To investigate the pathogenic mechanism of chronic wounds, we studied the effect of hypoxia on migration of keratinocytes isolated from human donors of increasing age. Keratinocytes from elderly donors had depressed migratory activity when exposed to hypoxia, as opposed to an increase in migration in young cells. Analysis of underlying biochemical changes demonstrated a differential activation of matrix metalloproteinases by hypoxia in keratinocytes isolated from the young and the old. Matrix metalloproteinases-1 and -9 and tissue inhibitor of matrix metalloproteinase-1 were strongly upregulated by hypoxia in young cells, whereas no induction was observed in aged cells. Furthermore, transforming growth factor-beta 1 signaling appears to be involved in the keratinocyte differential response to hypoxia, as transforming growth factor-beta type I receptor was upregulated by hypoxia in young cells, while there was no induction in aged cells. Transforming growth factor-beta neutralizing reagents blocked hypoxia-induced matrix metalloproteinase-1, matrix metalloproteinase-9 expression, and hypoxia-induced cell migration as well. Our results suggest that an age-related decrease in response to hypoxia plays a crucial part in the pathogenesis of retarded re-epithelialization in wound.

Transforming growth factor-beta. Effect on soft tissue repair.

Previous studies have demonstrated that TGF-beta possesses many of the biologic properties necessary for acceleration of the normal wound healing process. We report that recombinant human TGF-beta 2 (rhuTGF-beta 1) increases wound strength and accelerates wound closure when applied topically to experimental wounds. Doses of 5 to 1,000 ng/wound increased wound strength in a dose-response manner and wound strength increase as high as 161% above control in the rat incisional wound model. Increased wound strength was observed as early as 3 days following rhuTGF-beta 1 application and continued to Day 28. In the rabbit ear ulcer model, acceleration of wound closure was observed following doses of 5 to 100 ng/wound applied a single topical application. No adverse effects of rhuTGF-beta 1 were observed. The amount of fibrous tissue, scar formation, and mitotic figures were not significantly greater than control. Epithelialization of rhuTGF-beta 1-treated wounds was not impeded. rhuTGF-beta 1 induced bone formation in the rabbit ear ulcer model but not in the rat incisional model, suggesting that precursor cells, such as perichondrial cells, are required for the bone forming activities of TGF-beta 1.

Effect of ischemia on growth factor enhancement of incisional wound healing.

BACKGROUND: Several growth factors have been shown to enhance incisional wound healing in different models, but the models have been difficult to compare, and the effects under ischemic conditions are unknown. METHODS: An ischemic model was developed by selective interruption of the rabbit ear circulation and placement of dorsal incisions. The model was defined during a 28-day period by use of serial blood gas analysis, breaking strength measurement, and histologic analysis. In a separate experiment, incisions were treated with topical growth factors with the contralateral ear serving as paired controls, and the wounds were evaluated similarly. RESULTS: The ischemic ears were more hypoxic than controls through day 14 after wounding (48.5 versus 41 mm Hg, p < 0.03), and healing was impaired through day 28 (3.21 versus 1.90 newtons, p < or = 0.001). Transforming growth factor-beta (1 microgram) and Kaposi's fibroblast growth factor (20 micrograms) increased breaking strengths under both normal (3.03 versus 2.41 and 2.83 versus 2.47 newtons, respectively; p < 0.05) and ischemic conditions (1.40 versus 1.11 and 1.56 versus 1.23 newtons, respectively; p < 0.05). Platelet-derived growth factor-BB (10 micrograms) was effective only under normal conditions (2.67 versus 2.15 newtons, p < 0.02), whereas basic fibroblast growth factor (20 micrograms) was ineffective under both conditions. CONCLUSIONS: The rabbit ear incisional model is a reproducible ischemic incisional model allowing comparison of growth factor effects under ischemic and nonischemic conditions. Transforming growth factor-beta and Kaposi's fibroblast growth factor are both effective under ischemic conditions, whereas basic fibroblast growth factor and platelet-derived growth factor-BB are not, suggesting that ischemia is an important parameter affecting growth factor actions.

Platelet-activating factor: improvement in wound healing by a chemotactic factor.

Platelet-activating factor (PAF) is chemotactic for inflammatory cells and activates macrophages but, unlike growth factors, which have been demonstrated to accelerate healing, it has no effects on cell proliferation. The possible role of PAF in wound healing has not been explored previously. We examined the effect of different topical concentrations of PAF on the paired rat surgical incision model. Samples harvested on different days after wounding were evaluated for tensiometry and histology. Samples treated with 1 microgram, but not with 0.1 or 10 micrograms, showed an increase in maximal breaking strength (33.2%, 25.6%, and 4.9% by days 5, 7, and 12, respectively), reaching significance on days 5 and 7. Samples treated with 1 microgram PAF demonstrated a greater cellular infiltration (fibroblasts and monocytes) on day 7. Further histochemical studies revealed an increase of macrophages by day 7. Treatment with PAF receptor antagonist blocked the response to PAF but had no effect on normal wound healing, suggesting that, although PAF can accelerate healing, its endogenous production does not play a key role in normal wound repair. Our results demonstrate promotion of wound healing by PAF, a phospholipid chemotactic factor not previously shown to have wound-healing properties. This study gives new insights into how cytokines act to promote healing and suggests a strategy for improving wound repair with a monocyte chemotactic factor.

Topical silicone gel: a new treatment for hypertrophic scars.

A prospective, controlled clinical trial was designed to assess the efficacy of a new treatment of hypertrophic scars. Silicone gel sheeting was applied to 14 hypertrophic scars in 10 adults for 8 weeks. The treated scars and untreated, mirror-image or adjacent control scars were photographed, biopsy specimens were taken, and they were measured elastometrically before and after treatment. Photography and elastometry were repeated 4 weeks after treatment was discontinued. All the scars that had been treated for at least 12 hours a day were improved clinically after 4 weeks. There was further clinical improvement during the second 4 weeks of treatment. Elastometrically, the treated scars were improved significantly at 4, 8, and 12 weeks, compared with both their own treatment value and the control scars (p less than 0.05). Control scars were unchanged elastometrically. Clinical improvement persisted for at least 4 weeks after treatment was discontinued. The silicone gel sheeting was well tolerated, except for occasional transient rashes or superficial maceration--both of which resolved promptly when treatment was withdrawn. There was no histologic evidence of inflammation or foreign body reaction suggesting that silicone had entered the treated tissues. We conclude that this simple method of treating hypertrophic scar is efficacious, even in relatively chronic cases. The mechanism of action of silicone gel, which is apparently not related to compression, remains to be determined.

Granulocyte-macrophage colony-stimulating factor and granulocyte colony-stimulating factor: differential action on incisional wound healing.

BACKGROUND: Granulocyte-macrophage colony-stimulating factor (GM-CSF) specifically stimulates granulocyte, macrophage, and eosinophil colonies; granulocyte colony-stimulating factor (G-CSF) acts directly on neutrophil-restricted progenitor cells in their proliferation. Those cells have been implicated in the process of wound healing. METHODS: Paired 6 cm incisions were made on rats; GM-CSF or G-CSF was given systemically (100 micrograms/kg/dose) or locally (30 micrograms/wound). The controls received vehicle alone. Impaired healing was induced by injection of methylprednisolone (30 mg/kg). White blood cells (WBC) were counted at day 2 after treatment. Tissue strips were evaluated for tensiometry and histologic features at days 7 and 14 after wounding. RESULTS: For local GM-CSF treated incisions, the breaking strength was 25% stronger than controls at day 7 (p = 0.004), 36% at day 14 (p < 0.0001), and 42% at day 7 (p = 0.012) in impaired animals. Local G-CSF and systemic GM-CSF and G-CSF increased circulating WBC (p < 0.05), but they had no effects on healing. Histologic studies revealed an increase of wound cellulity at day 7 and day 14 in topical GM-CSF treated wounds. CONCLUSIONS: These results suggest GM-CSF is an activator of tissue macrophages and that increasing circulating WBC did not affect wound healing.

Acceleration of tissue repair by transforming growth factor beta 1: identification of in vivo mechanism of action with radiotherapy-induced specific healing deficits.

BACKGROUND: Transforming growth factor beta 1 (TGF-beta 1) is an effective accelerator of soft tissue repair in both normal and impaired healing models; however, its in vivo mechanism of action remains unclear. Modern radiation techniques can create unique healing deficits, allowing for a more specific definition of tissue response to growth factor therapy. In the rat linear skin incision model, cobalt 60 photon beam total body irradiation (TBI), 800 rads, causes a marked depression of circulating monocytes and largely spares the skin tissue. Megavoltage electron beam surface irradiation (SI), 2500 rads, markedly impairs surface healing while sparing the bone marrow. With these models of selective healing deficits, the ability of TGF-beta 1 to accelerate tissue repair directly in the absence of circulating macrophage precursors (TBI) or in the presence of damaged dermal fibroblasts (SI) was evaluated. METHODS: Adult male Sprague-Dawley rats were randomly assigned to groups of TBI, SI, or nonirradiated sham controls and received radiation 2 days before wounding. Paired linear full-thickness skin incisions were created and a single dose of TGF-beta 1 (2 micrograms/wound) or vehicle control was applied to each wound. RESULTS: Both radiation techniques produced a marked healing deficit when assessed on postwounding days 7 and 12. TBI treatment was characterized by severe monocytopenia, confirmed by a tissue macrophage-specific immunohistochemical technique. On days 7 and 12 after wounding, TGF-beta 1 significantly accelerated soft tissue repair and wound-breaking strength in the TBI-treated rats, demonstrating an ability to directly promote the induction of collagen synthesis in the absence of monocytes/macrophages. In contrast, TGF-beta 1 was unable to reverse the SI-induced healing deficit characterized by impaired function of dermal fibroblasts. CONCLUSIONS: These in vivo observations provide further evidence for a direct mechanism of action by TGF-beta 1 on collagen synthesis by wound fibroblasts during soft tissue repair.

Differential acceleration of healing of surgical incisions in the rabbit gastrointestinal tract by platelet-derived growth factor and transforming growth factor, type beta.

Anastomotic dehiscence is a major cause of morbidity and mortality in gastrointestinal surgery. A unique model system of a gastric incision was developed to test the potential of polypeptide growth factors to enhance wound healing. Paired, deep partial-thickness incisions to but not including the gastric mucosa were made. A single topical application of transforming growth factor, type beta 1 (TGF-beta), platelet-derived growth factor, or control vehicle at the time of wounding was given. Wound breaking strength and detailed histologic analyses of wounds were evaluated as a function of time after wounding. TGF-beta (0.1 to 2.0 micrograms/wound) demonstrated a bimodal, dose-dependent acceleration of wound breaking strength 7 days after gastric wounding. An approximate 4-day acceleration of gastric wound breaking strength by TGF-beta (2 micrograms/wound) was seen at 7 and 11 days. Wounds treated with platelet-derived growth factor (10 micrograms/wound) displayed an increased cellular response but no enhancement of breaking strength at 7 and 11 days. These results demonstrate the ability of TGF-beta to accelerate gastrointestinal tissue repair by topical application and suggest significant potential for the use of growth factors in enhancing repair of surgical wounds of the gastrointestinal tract.

The effect of new technologies on plastic surgery.

Plastic surgery has always been a technique- and technology-driven surgical discipline, given that there is no regional anatomic focus. There has been a remarkable evolution in technique over the last 25 years with an increased understanding of anatomy leading to a whole host of new and more reliable flaps, which has transformed reconstructive surgery, breast reconstruction being one notable example. The development and maturation of microsurgery has led to the full fruition of anatomic principles. With better understanding of blood supply to the skin, fascia, muscle, and bone, many traditional reconstructive procedures are constantly being superseded by the new, ingenious use of various tissue flaps. Advances in technology will accomplish another transformation of the specialty, notably the recent advances in tissue engineering, the potential of gene therapy, new alloplastic materials, and computer-assisted imaging technology. It would be impossible to address all of the recent advances in this rapidly expanding field of surgery in a short article. We have selected a few topics that we thought would be the most interesting to all surgeons to give a wide view of a variety of challenges addressed by the modern plastic surgeon. Major advances in surgery often come from cross-fertilization between specialties, and plastic surgeons have frequently been involved in this process.

Topical silicone gel for the prevention and treatment of hypertrophic scar.

We studied the effects of a silicone gel bandage that was worn for at least 12 hours daily on the resolution of hypertrophic burn scar. In a second cohort, the prevention of hypertrophic scar formation in fresh surgical incisions by this bandage was also evaluated. In 19 patients with hypertrophic burn scars, elasticity of the scars was quantitated serially with the use of an elastometer. An adjacent or mirror-image hypertrophic burn scar served as a control. Scar elasticity was increased after both 1 and 2 months compared with that in controls. There was corresponding improvement clinically that persisted for at least 6 months. In the other cohort, scar volume changes in 21 surgical incisions were measured before and after 1 and 2 months. Gel-treated incisions gained less volume than control incisions after both intervals. Clinical assessment corroborated this quantitative demonstration of a decrement in scar volume. We concluded that topical silicone gel is efficacious, both in the prevention and in the treatment of hypertrophic scar.

Keratinocyte growth factor induces granulation tissue in ischemic dermal wounds. Importance of epithelial-mesenchymal cell interactions.

BACKGROUND: Keratinocyte growth factor acts specifically on epithelial cells and is presumed to play an important role in tissue repair. OBJECTIVE: To examine the wound-healing effects of keratinocyte growth factor under hypoxic conditions in vivo and in vitro. DESIGN AND INTERVENTIONS: Dermal ulcers were created in the ischemic ears of 40 anesthetized young female rabbits. Either recombinant keratinocyte growth factor (rKGF) or buffer was applied to each wound. Wounds were bisected and analyzed histologically at days 7 and 10 after wounds were created. For the in vitro study, normal keratinocytes were treated with rKGF (20 ng/mL) and cultured under hypoxic (3.5% oxygen) conditions. The conditioned media were collected at 48 and 72 hours. MAIN OUTCOME MEASUREMENTS: The amount of epithelial growth and deposition of granulation tissue were measured in all wounds. The amount of transforming growth factor alpha in keratinocyte-conditioned media was measured by using a sensitive radioimmunoassay. A proliferation assay of dermal fibroblasts, treated with conditioned media, was also performed under 3.5% oxygen culture conditions. RESULTS: The rKGF (range, 5-40 micrograms per wound) that was applied significantly increased new epithelium by greater than 70% (P = .03) at days 7 and 10 after wounds were created. A significant increase in new granulation tissue formation (170%) was also observed in rKGF-treated wounds at day 10, at a dose of 40 micrograms per wound (P < .002). The amount of transforming growth factor alpha protein in the conditioned media that were treated with rKGF (20 ng/mL) increased by 26.8% and 171% at 48 and 72 hours, respectively, over that of controls. The conditioned media from rKGF-treated keratinocytes, grown for 72 hours, resulted in a 51% increase in the proliferation of primary rabbit dermal fibroblasts. CONCLUSION: Keratinocyte growth factor enhances the wound-healing process of ischemic ulcers, indicating that epithelial-mesenchymal cell interactions are critical for the healing of wounds under ischemic conditions and possibly under normal conditions as well.

Effect of hyperbaric oxygen and growth factors on rabbit ear ischemic ulcers.

OBJECTIVE: To test the influence of hyperbaric oxygen (HBO), platelet-derived growth factor-BB (PDGF-BB), and transforming growth factor-beta 1 (TGF-beta 1) on the deficit in wound healing produced by ischemia in a noncontractive dermal ulcer standardized model in the rabbit ear. DESIGN AND INTERVENTIONS: Dermal ulcers were created in the ischemic ears of 42 anesthetized young female New Zealand white rabbits. The controls were ulcers created in nonischemic ears of eight anesthetized young female New Zealand white rabbits. Either PDGF-BB (5 micrograms), TGF-beta 1 (1 microgram), or buffer alone was applied to each wound, which was then covered. Some groups were treated with HBO on days 0 through 4. Wounds were harvested on day 7 and were evaluated histologically. MAIN OUTCOME MEASURES: The amount of epithelial regrowth and granulation tissue production were measured. The wounds were evaluated for glycosaminoglycan and collagen content. Angiogenesis was measured. RESULTS: Hyperbaric oxygen alone, in the ischemic model, increased the production of new granulation tissue by approximately 100% at 7 days without significantly affecting new epithelial growth (P = .03). In contrast, PDGF-BB and TGF-beta 1 each increased the new granulation tissue volume by greater than 200% in 7 days (P = .0001) and also had a statistically significant effect on new epithelial growth. However, the addition of growth factors to HBO treatment produced a synergistic total reversal of the wound-healing deficit produced by ischemia (P = .0001). CONCLUSIONS: Both PDGF-BB and TGF-beta 1 alone are more effective than HBO treatment by itself in accelerating the impaired wound healing produced by ischemia. However, the combination of HBO with either of the growth factors has a synergistic effect that totally reverses the deficit produced by ischemia.

Transforming growth factor beta 3 (TGF beta 3) accelerates wound healing without alteration of scar prominence. Histologic and competitive reverse-transcription-polymerase chain reaction studies.

BACKGROUND: Transforming growth factor (TGF) beta 3 is a new isoform of the TGF beta superfamily and is presumed to play an important role in wound repair and scarring. OBJECTIVE: To examine the effects of TGF beta 3 on wound healing and on reducing scarring. DESIGN AND INTERVENTIONS: Dermal ulcers were created on the ears of 75 anesthetized young female rabbits. Either TGF beta 3 or vehicle was applied topically to the wounds. Wounds were bisected and analyzed histologically at postwounding day 7. A second group of wounds was treated with topical TGF beta 3 and TGF beta 2 or vehicle at days 0 and 3 and harvested at days 21 through 42 as an excessive scarring model. The third group of wounds was treated with TGF beta 1, TGF beta 2, and TGF beta 3 and vehicle. The granulation tissue was harvested at day 7, and cellular RNA was extracted for performing competitive reverse-transcription polymerase chain reaction. MAIN OUTCOME MEASUREMENT: The amount of new epithelium and granulation tissue was measured in TGF beta 3- and vehicle-treated wounds. The hypertrophic index was calculated for scarring wounds treated with TGF beta 2 and TGF beta 3 or vehicle. Levels of TGF beta 1 messenger RNA were measured in those wounds that were treated with TGF beta 1, TGF beta 2, and TGF beta 3 and in their controls. RESULTS: The use of TGF beta 3 (0.3-0.75 microgram per wound) increased granulation tissue formation by more than 100% (P < .005). Epithelialization showed a biphase, either increasing 30% (P < .04) or decreasing 25% (P < .001) dependent on dose. No significant difference in the hypertrophic index was noted in TGF beta 3-treated wounds compared with controls. Levels of TGF beta 1 messenger RNA increased (7.1- to 14.9-fold) in those wounds treated with TGF beta s compared with controls at day 7. CONCLUSIONS: Exogenous TGF beta 3 displays substantial vulnerary properties in wound healing and may be useful in treating nonhealing wounds. However, the observation that TGF beta 3 can reduce scarring was not confirmed in this study, and the messenger RNA level in response to TGF beta 3 suggests that it behaves similarly to TGF beta 1.

Enhancement of wound healing by hyperbaric oxygen and transforming growth factor beta3 in a new chronic wound model in aged rabbits.

HYPOTHESIS: Although hyperbaric oxygen (HBO) has been used clinically for 3 decades, there have been few controlled clinical trials. Animal models have not been adequate to test the efficacy of HBO in the treatment of chronic wounds, either by itself or in combination with growth factors. We hypothesize that HBO is as efficacious as a prototype growth factor in improving wound healing in a new animal model of ischemic chronic wounds. DESIGN: Twenty-five aged rabbits and 3 young rabbits had their ears rendered chronically ischemic and ulcers were created down to the level of cartilage. These ulcers were treated in 1 of 3 ways: with HBO, 90 minutes per day, Monday through Friday, for 4 weeks; with transforming growth factor beta(3) at 1 microg/cm(2); or with both modalities combined. Controls were treated with vehicle or hyperbaric room air or both. RESULTS: This model created an aged/ischemic wound that failed to heal spontaneously up to 26 days after wounding (88% reduction compared with aged/nonischemic controls). Hyperbaric oxygen alone and transforming growth factor beta(3) alone both improved healing rate (only 38% reduction in healing compared with aged/nonischemic controls). Combined therapy produced no additional improvement over either modality by itself. CONCLUSIONS: In aged animals, HBO and transforming growth factor beta(3) were equally effective in improving wound healing. Our data suggest that HBO alone may be more effective in the chronic wound than in the acute wound. There was no additive benefit to combining modalities as has been reported in the same wound model in young rabbits.