ABSTRACT
The tissue reaction of pig skin to implantation of decellularized and recellularized dermal matrices on a formed wound defect was evaluated by histological methods on days 2, 5, 8, 16, and 20 after surgery. Differences in tissue response to different matrices were identified. In experimental wounds coated with decellularized dermal matrices, we observed the formation of a scar tissue, which required autodermoplasty on day 12 of the experiment. In wounds coated with recellularized dermal matrices, all layers of the skin completely recovered by day 20 after surgery with the formation of full dermal and epidermal layers. Our findings suggest that reparative morphological changes in the wound depend on the presence of fibroblasts in the implanted dermal matrix.
Subject(s)
Skin/cytology , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Animals , Extracellular Matrix , Fibroblasts/cytology , Swine , Wound Healing/physiologyABSTRACT
Automated image analysis methods are highly important for biotechnology research. The authors developed and tested a program for the morphometric analysis of photomicrographs of the sections processed using the standard immunohistochemical examination protocols. The color deconvolution method used in the algorithm was proven to be effective in mapping the distribution of DAB chromogen in the sample containing multiple dyes. The experiment demonstrated that the level of extracellular matrix proteins could be comparatively quantified in different groups of samples. The effective methods for the quantitative analysis of the Ki-67 labelling index were also tested using the same algorithms. The developed program was published under free GPL 3.0.
Subject(s)
Antigens, Nuclear/isolation & purification , Extracellular Matrix/ultrastructure , Image Processing, Computer-Assisted/methods , Immunohistochemistry/methods , Algorithms , Antigens, Nuclear/ultrastructure , Coloring Agents/chemistry , Coloring Agents/isolation & purification , Humans , SoftwareABSTRACT
AIM: To develop pathogenetically justified surgical treatment of diabetic neuroosteoarthropathy (DNOAP). MATERIAL AND METHODS: 52 patients were operated and long-term results were studied in 36 of them. 2 groups were assessed depending on completeness of surgical treatment including changed synovial structures removal. Tarso-metatarsal arthrodesis was created after synovectomy in median foot to restore longitudinal arch. Cruro-calcaneal neoarthrosis was performed after elimination of posterior focus of DNOAP. Excision of large, deep plantar ulcer (or osteomyelitic fistula) was completed by tissues tension with needles for primary wound healing. RESULTS: DNOAP patients are tolerant against surgical infection that is decreased in patients with neuropathic form of diabetic foot syndrome. Skin plasty is followed by rejection in DNOAP that was cured with prednisolone. Deep suppurations complicating foot skeleton fragmentation have subacute course and do not lead to sepsis. Infected wounds heal by primary intention in 98% after closure with primary suture. It was empirically found that synovectomy and osteochondral detritus removal discontinue DNOAP course. Reconstructive surgical stage is aimed to restoring the shape and function of the foot. Analysis of long-term outcomes showed absence of DNOAP signs in 76.2±9.2% of cases after radical surgery and positive results of treatment in 97.2±2.7% of patients. CONCLUSION: Our data conceptually assume the possibility of autoimmune mechanism in DNOAP pathogenesis. On this basis pathogenetically reasonable surgical procedures are established to interrupt pathological process. These interventions differ by technological aspects only depending on middle or rear foot destruction.