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Skin development in bony fish with particular emphasis on collagen deposition in the dermis of the zebrafish (Danio rerio).

The first part of this article is a review of the current status of knowledge of the fish skin, with particular attention to its development. In the second part we present original results obtained in zebrafish (Danio rerio), with particular emphasis on the deposition and organisation of the dermal collagenous stroma. Using a series of zebrafish specimens aged between 15 hours postfertilization (hpf) and 4.5 years old, we have combined Transmission Electron Microscopy (TEM) observations and in situ hybridisation using type I collagen a2 chain (Col1a2) probe. Collagen fibrils, with a diameter of 22 nm, appear first in an acellular subepidermal space at 24 hpf, are first all oriented in the same direction, and form the primary dermal stroma. Subsequently, three events occur. (1) From 5-7 days pf (dpf) onwards the collagen fibrils self-organise into several lamellae arranged in a plywood-like structure, starting in the upper layers and progressing throughout the entire thickness of the dermis. (2) At 20-26 dpf, fibroblasts of unknown origin progressively invade the acellular collagenous stroma, some of them accumulating below the epidermis. (3) Concomitant with the invasion of fibroblasts, the collagen fibrils increase progressively in diameter to reach 160 nm towards the end of the fish life. In situ hybridisation experiments reveal that, between 24 and 48 hpf, the collagen matrix is produced by the epidermis only. From 72 hpf to 20-26 dpf, both the basal epidermal cells and the dermal cells bordering the deep region of the dermis are involved in the production of collagen. When the fibroblasts invade the plywood-like structure, the epidermal cells progressively cease to synthesise collagen, which from this point is produced only by the fibroblasts. This suggests that the fibroblasts secrete a still unidentified signalling molecule that downregulates collagen production by the epidermis.

Animals↗

Study of gelatin-containing artificial skin V: fabrication of gelatin scaffolds using a salt-leaching method.

Porous gelatin scaffolds were prepared using a salt-leaching method and these were compared to scaffolds fabricated using a freeze-drying method. The salt-leached gelatin scaffolds were easily formed into desired shapes with a uniformly distributed and interconnected pore structure with an average pore size of around 350 microm. The mechanical strength and the biodegradation rate of the scaffolds increased with the porosity, and were easily modulated by the addition of salt. After 1 week of in vitro culturing, the fibroblasts in salt-leached scaffolds were mainly attached on the surface of the pores in the scaffold, whereas cells seeded on freeze-dried scaffolds were widely distributed and aggregated on the top and the bottom of the scaffold. After 14 d of culturing, the fibroblasts showed a good affinity to, and proliferation on, the gelatin scaffolds without showing any signs of biodegradation. An in vivo study of cultured artificial dermal substitutes showed that an artificial dermis containing the fibroblasts enhanced the re-epithelialization of a full-thickness skin defect when compared to an acellular scaffold after 1 week.

Animals↗

Human leucocyte migration: studies with an improved skin chamber technique.

An improved skin chamber technique has been developed for the study of localized leucocyte mobilization (LLM). Uniform "windows" of denuded dermis were produced by a suction device applied to the forearm skin, eliciting delineated areas of epidermal separation by blister formation. The acellular blister fluid, roof and basement membrane were removed, and the blister base was covered with a rubber chamber containing autologous serum as leucocyte attractant. Duplicate chambers were harvested at prescribed intervals during the first 24 hours. In 15 healthy individuals, virtually no cells were observed after 2 hours, a median of 1.9 X 10(6) after 4 hours, increasing to 3.8 X 10(7) after 24 hours. Subnormal LLM was demonstrated in three of seven patients with severe bacterial infections and in three of seven leukaemia patients. LLM was normal in eight patients with other malignancies. Ninety to 98 per cent of the cells were polymorphonuclear neutrophils and less than 1 per cent were erythrocytes. In the chamber neutrophils, vacuolization of the cytoplasm was prominent, bactericidal capacity reduced and nitroblue tetrazolium reduction increased, thus indicating functional derangement of emigrated cells compared to peripheral blood neutrophils. Simplicity and good reproducibility should make this method a valuable tool in the study of leucocyte migration.

Adolescent↗

The use of a permanent dermal allograft in full-thickness burns of the hand and foot: a report of three cases.

The standard method of grafting deep, thermal hand-and-foot burns with either full-thickness sheet grafts or narrowly meshed, thick, split-thickness skin grafts not only leaves a deep donor site, but also becomes complicated by infection, hypertrophic scarring, blistering, and hyper- or hypopigmentation. The availability now of an acellular, immunologically inert dermal transplant (AlloDerm; LifeCell Corp., The Woodlands, Texas) allows the successful use of ultrathin autografts while maximizing the amount of dermis delivered to the wound site. These autografts leave thin donor sites that heal faster and with fewer complications. This case report describes the use of AlloDerm dermal grafts on three patients with full-thickness burns of the distal extremities. Grafts were applied to the hand in two cases and the dorsum of the foot in the third. Range of motion, grip strength, fine motor coordination, and functional performance were quantitatively evaluated. As demonstrated by these patients, cosmetic and functional results were considered good to excellent after the use of AlloDerm grafts with thin autografts.

Adult↗

Use of allogenic dermis for radial forearm free flap donor site coverage.

OBJECTIVE: The radial forearm free flap has become the method of choice for reconstruction of head and neck defects following oncologic ablation. Harvesting of a radial forearm free flap leaves a donor site defect. This is most commonly closed with a split-thickness skin graft. Morbidity, most commonly owing to a lack of graft take over the tendons, can be quite high. Recently, an acellular matrix (Alloderm) has been advocated to decrease complications at the radial forearm donor site, as well as obviate taking a split-thickness skin graft from the thigh. MATERIAL AND METHODS: Tertiary referral academic centre. Retrospective chart review of 15 patients. Five patients received allogenic dermis, 10 patients received split-thickness skin grafting to the radial forearm donor site. RESULTS: Patients with allogenic dermis took between 12 and 16 weeks to heal completely. Patients undergoing split-thickness skin graft were completely healed within 4 to 6 weeks. Cosmesis was judged to be marginally better in the allogenic dermis group. Allogenic dermis placement had a greater impact on hand function owing to prolonged healing, whereas patients with split-thickness skin graft required wound care at the thigh for a 2- to 3-week period owing to the harvesting of the skin graft. CONCLUSIONS: Allogenic dermis may be a viable alternative to split-thickness skin grafting and radial forearm free flap donor sites. Prolonged healing with subsequent increased health care services use needs to be addressed.

Aged↗

Evaluation of a porcine origin acellular dermal matrix and small intestinal submucosa as dermal replacements in preventing secondary skin graft contraction.

The degree to which a split thickness skin graft (STSG) contracts after application to its recipient bed is related in part to the proportion of the dermis harvested from the donor site. Harvesting thicker skin grafts may produce better cosmetic results in the recipient bed but result in increased donor site morbidity. The combination of an autologous ultra thin split thickness graft with an underlying non-autologous dermal component may reduce secondary skin graft contraction without further increasing donor site morbidity. This study was aimed at assessing the suitability of two porcine derived biomaterials (Permacol and small intestinal submucosa, SIS) for use in combination with skin grafts in a Sprague-Dawley rat model. Full thickness wounds (1 cm(2)) were created in Sprague-Dawley rats and grafted with skin in combination with Permacol or SIS either as a one-stage operation or following a 2-week-period of vascularisation of these dermal matrices before a second stage operation to cover with skin. Skin graft viability and wound area were assessed at weekly intervals until 4 weeks after graft application. Both Permacol and SIS were able to support an overlying skin graft but had no beneficial effect on skin graft contraction in this model compared to skin grafts alone.

Animals↗

Dispase/detergent treated dermal matrix as a dermal substitute.

A method for preparing acellular allogeneic dermal matrix (ADM) and its effectiveness as a dermal substitute are described. Treatment of rat skin with Dispase followed by Triton X-100 completely removed cellular components from the dermis. Subcutaneously implanted ADM evoked no immunological reaction and 20 weeks after implantation, the size of the implanted ADM was reduced to about 60 per cent of its original area. ADM became completely vascularized within 2 weeks after implantation into full thickness skin defects in the rat and inhibited extensive wound contracture. A second layer of ADM placed onto the implanted ADM served as an excellent dressing, providing mechanical protection and permitting vascularization of the underlying implant. Onlay skin autografts placed onto vascularized allogeneic ADM showed good survival when the skin was grafted more than 1 week after ADM implantation. Dispase/detergent treated ADM derived from animal or human skin may be useful in full thickness skin defects providing a vascularized bed for subsequent epidermal coverage.

Animals↗

Creation of an acellular dermal matrix from frozen skin.

At present, one of the treatments of choice for closure of full-thickness skin loss is to use a cultured epidermal autograft when skin loss is extensive. In this study, we investigated a simple method of processing frozen surplus skin to produce an acellular, structurally intact, dermal matrix. First, the acellular dermal matrix prepared from normal human skin (ADM) we processed was observed using a transmission electron microscope and a scanning electron microscope. The matrix maintained the basement membrane complex and the extracellular matrix structure of the dermis despite frozen skin being used. Next, using an animal model, we transplanted the ADM and Pelnac, which is used as a contrast in full-thickness wounds onto nude rats. The dermal matrix supported fibroblast infiltration and neovascularization. These results suggest that skin processed by our simple method has the potential to be used as a dermal template together with the cultured epidermis in the closure of full-thickness wounds.

Animals↗

[The clinical application of acellular dermal matrix to repair the defect of oral mucosa].

OBJECTIVE: To evaluate the clinical effect of acellular dermal matrix allograft in repairing the defect of oral mucosa. METHODS: Forty-two cases of large superficial defects in oral cavity were repaired by acellular dermal matrix. Defect areas included vestibule, tongue, the floor of mouth, buccal mucosa, palate, gingival and lip. Defect size was between 2.25 cm(2) and 12 cm(2). The basement membrane side of acellular dermal matrix was exposed to oral cavity, and another side was attached to the surgical wound surface. It was fixed by suture, reverse strapping and prepared template to overlaid large superficial defect of oral mucosa. The patients were followed up from 3 months to 1 year after operation. RESULTS: The wounds were completely healed in 40 cases, partially healed in 2 cases. There was no failure case. The healing was divided into two types: epithelialization healing pattern in 12 cases, attached healing pattern in 30 cases. Contraction of grafts occurred at 2 - 4 weeks. The contraction rate was 3.7% +/- 1.1% at fourth week. Scarring did not appear. CONCLUSION: The clinical result of acellular dermal matrix allograft in repair the defect of oral mucosa is good. Acellular dermal matrix can be taken as a substitute to autograft of skin.

Adult↗

Gingival augmentation with an acellular dermal matrix: human histologic evaluation of a case--placement of the graft on bone.

The importance of gingival augmentation to increase the amount of keratinized tissue is a controversial subject. In this case report, an acellular dermal matrix was used to obtain an increase in the amount of keratinized tissue around four implants. In this case, the acellular dermal matrix was placed on bone. The surgical procedure resulted in an increase in the amount of keratinized tissue. Therefore, it met the clinical goals of the surgical procedure. However, the clinical findings and patient pain levels during the healing seemed to resemble a denudation procedure. Additionally, the histologic evaluation of the tissue that formed around the implants showed that the acellular dermal matrix was not incorporated into the result. Based on this case, the use of an acellular dermal matrix placed on bone does not seem to be a good technique to increase the amount of keratinized tissue.

Aged↗

[The clinical application of xenogenic (porcine) acellular dermal matrix grafting with thin autogenic skin on deep wounds].

OBJECTIVE: To observe the effectiveness of xenogenic(porcine) acellular dermal matrix grafting with thin autogenic skin on deep wounds with good wound-bed. METHODS: 52 cases of patients with deep wounds were transplanted with xenogenic(porcine) acellular dermal matrix grafting and thin autogenic skin by two step method from Jan. 2000 to Feb. 2002. RESULTS: 37 cases of patients achieved complete take(71.1%) and 15 cases got 95 percent take (28.9%) with none needing grafting again. 3-6 months' following up showed that skins of complex grafting had well-acceptable color and quality compared to nearby skin. CONCLUSION: This kind of xenogenic(porcine) acellular dermal matrix can be widely used on deep wounds with good wound bed as complex graft.

Adolescent↗

[Experimental study and clinical application of composite skin grafting].

OBJECTIVE: To explore an ideal skin substitute with its appearance and texture similar to normal skin, to repair wounds with full-thickness skin defect. METHODS: Composite skin (CS) in question was composed of allo/xenogeneic acellular dermal matrix (ADM) and razor thin autoskin. One step skin grafting was employed in the experimental study and clinical trial. Razor thin autoskin alone was used as the control in the study. Changes in the antigenicity of ADM and the reformation of basement membrane (BM) structure at epidermis-dermis junction (EDJ) of ADMs were studied at designated time points after the grafting with biochemical and immunohistochemical methods. Fifty-three patients with full thickness skin defects due to various causes, including scar excision were grafted with CS, and survival rate and long-term result were observed. RESULTS: The grafted CS survived satisfactory. The reformation of the basement membrane structure was clearly observed at the 28th post-graft week. The basement membrane cells grew with polarization in an undulating arrangement. There was reformation of dermal papillae and ridges. The antigenicity of allo-ADM was obviously lower than that of xeno-ADM. Sixty-five out of 70 pieces of CS grafting (92.9%) survived totally, two of them survived partially, and three failed due to infection. The longest follow-up period was 8 and a half years. The grafted CS appeared similar to the normal skin in regard to the texture and color, especially allo-ADM, and no evident rejection reaction was seen. CONCLUSION: ADM possessed very low antigenicity, thus serving a lasting framework after grafting. In addition, it could serve as a "dermal template" for the induction of tissue regeneration.

Animals↗

[Preliminary study on outcome of xenogeneic (porcine) acellular dermal matrix in vivo].

OBJECTIVE: To investigate the outcome of xenogeneic (porcine) acellular dermal matrix in vivo. METHODS: Xenogeneic (porcine) acellular dermal matrix was prepared by removing cellular components with trypsin and triton X-100 and was embedded under the skin of SD rat. Gross and histological changes were observed periodically. RESULTS: There was no cellular components but with intact basal membrane, and collagen was found to have a regular arrangement in xenogeneic (porcine) acellular dermal matrix. Grossly, the xenogeneic (porcine) acellular dermal matrix was found to disintegrate gradually. Histological examination revealed that xenogeneic (porcine) acellular dermal matrix was mainly infiltrated by inflammatory cells in one week after implantation. After one week, fibroblasts and capillary vessel increased in number gradually, and collagen began to arrange densely in regular manner. CONCLUSION: Xenogeneic (porcine) acellular dermal matrix disintegrates gradually after transplantation, inducing growth of fibroblasts and capillaries.

Animals↗

An in vitro examination of an extracellular matrix scaffold for use in wound healing.

This paper describes evidence that an extracellular matrix (ECM) secreted by human umbilical vein endothelial cells (HUVECs) assembled on gelatin coated plates overlaid by a mixed matrix secreted by human dermal microvascular endothelial cells (HDMECs) and human dermal fibroblasts provides a viable acellular scaffold for use in wound healing. Trypsinized epidermal keratinocytes or colonies from Dispase-digested fresh and cadaver skin tissue adhered and proliferated on either HUVECs ECM/gelatin or mixed matrix overlaid on HUVECs ECM/gelatin. An epithelial-mesenchymal interaction, previously thought to be tissue-specific, was exposed as well as concomitant integrin versatility. Furthermore, heterologous HDMECs and dermal fibroblasts attached and proliferated on the mixed matrix as well as HUVECs ECM. The conditioned medium from HUVECs (HUVECs CM) was found to neutralize the lingering after effects of Dispase, and could be used for the tissue culture of epidermal keratinocytes, HDMECs and dermal fibroblasts, which share related extracellular secretions. Taken together, these results indicate that cultured epithelial autografts can be redesigned to include both epithelial and dermal elements, and advances the acellular 'sandwich' ECM scaffold as a possible structural replacement for the lamina densa and lamina lucida, damaged or completely missing in some wounds and burns.

Burns↗

[Experimental study on the inflammatory and immune responses of xenogenic acellular dermal matrix transplantation combined with thin split-thickness skin autograft].

OBJECTIVE: To investigate the dynamic process of the inflammatory response and the profile of Th1/Th2 cytokines after xenogenic acellular dermal matrix (ADM) transplantation with thin split-thickness skin autograft overlay. METHODS: SD rats were used in the study. In the control group, thin split-thickness skin autograft (STSG) was transplanted in the full-thickness skin defect of the SD rats; in the experimental group, the xenogenic acellular dermal matrix combined with thin split-thickness skin autograft was transplanted. The inflammatory response was examined histologically and Th1/Th2 cytokine mRNA expression in skin grafts was determined by reverse transcription-polymerase chain reaction. RESULTS: Inflammatory reaction was induced by ADM at the early stage of transplantation and decreased gradually. Th2 cytokine mRNA expression was higher in the ADM group than that of the control group whereas the Th1 cytokine mRNA expression was undetected in both groups. CONCLUSION: Xenogenic acellular dermal matrix is immunogenic. The increased expression of Th2 cytokines may be related to the humoral immune responses and the absence of ADM graft rejection.

Animals↗

[Homologous dermal acellular matrix graft for urethral reconstruction in man (report of 16 cases)].

OBJECTIVE: To investigated the value of homologous dermal acellular matrix graft (AMG) for urethral reconstruction in man, and found an ideal replacement material to simplify the urethroplastic surgery. METHODS: 16 male patients, aged 18-46 (mean 26.5), with urethral diseases were treated with allograft of tissue engineering material AMG. The causes of urethroplasty were complex urethral stricture followed by pelvic fracture (13 cases), anterior urethral stricture (2 cases), hypospadia (1 case). During the operation, AMG was sutured to a tubular graft and replaced the defect urethra. A 18-22 Fr silicone catheter was inserted in the reconstructed urethra for 4-6 weeks. RESULTS: All 16 patients were voiding well after removing the catheter, urethrography reveal excellent caliber of the reconstructed urethra. Urethroscopic examination showed the graft urethra was covered by epithelial tissue and grew into the native tissue. But stenosis was developed in 3 patients postoperatively, 1 patients received transurethral incision, another 2 needed urethral sounding. The patients were followed-up for 12-72 months (mean 45.6), no rejection was observed, all patients voided normally except 4 cases needed urethral dilatation periodically. CONCLUSION: The homologous dermal acellular matrix graft may serve as an ideal replacement material for complex urethral stricture or defect, without concern of rejection.

Adolescent↗

Onlay hiatal reinforcement utilizing human acellular dermal matrix: three case series.

Redo laparoscopic fundoplication and laparoscopic repair of large (>5cm) paraesophageal hernias have a high rate of recurrence after primary suture repair of the hiatal defect. As such, the use of mesh prosthesis as an interposition graft or onlay reinforcement is becoming more popular for the repair of larger, more complicated crural defects. We report three cases in which human acellular dermal matrix was used as an onlay reinforcement of the hiatus after primary suture closure. Two patients had large paraesophageal hernias (one type III and one type IV). The third patient became symptomatic after her second laparoscopic antireflux procedure and was found to have recurrent herniation of the fundus into the mediastinum. All three patients underwent successful laparoscopic repair. There were no intraoperative or postoperative complications. All three patients remain symptom free with follow-up ranging from 8 months to 10 months. Acellular dermal matrix appears to be a promising prosthetic for onlay reinforcement of the hiatus during redo laparoscopic fundoplication and repair of large paraesophageal hernias.

Aged↗