# Evaluation of Keratin/Bacterial Cellulose Based Scaffolds as Potential Burned Wound Dressing
## 3 Results and Disscusion
### In Vivo Tests  
For the clinical testing of the scaffolds, eight common breed rabbits were included. A general clinical examination was performed before anesthesia and included the assessment of the habitus, skin and mucous membranes, the examination being repeated and post-intervention. The results also depend on various co-morbidities, in multicellular organisms, both health and disease are defined by means of communication patterns involving the component cells. Clinical supervision was performed daily throughout the experiment; no changes in the physiological state were reported in the rabbits included in the study [54; 55; 56; 57]. The appearance of the wound was followed and a favorable evolution was observed after the graft. One noticed the obvious recurrence of hairs throughout the wound contour, the absence of inflammation, and after taking biological samples for anatomic-pathological tests, the revascularization of the tissue [4; 58]. Palpation of the area shows a hard, normal appearance, without a painful sensation. The evolution of wound recovery is presented in the next images (Figure 8), where, M is the wound grafted with biomaterial made of BC-\(\beta\)-keratose and X wound grafted with biomaterial of BC-\(\beta\)-keratose and artificial tegument. Histopathological evaluation was carried out on the marginal areas of wounds, in all experimental groups. The morphology of the local inflammation, the conjunctive-vascular buds' formation and the epidermis regeneration process were monitored. At seven days after grafting the histopathological exam (after the established criteria) did not reveal any significant differences between the animal groups (Figure 9). All the studied animals developed increased dermo-epidermal coagulation necrosis, inflammatory infiltration in the hypoderm, represented by neutrophilic leukocytes, histiocytes and macrophages, all implicated in necrotic tissue detorsion. A congestion of the blood capillaries and a pronounced local edema, with the dissociation of local structural elements, was observed in the hypoderm. Figure 7: Cell viability data for BC-keratose scaffolds (MTT assay at 24 h, 48 h, and 72 h). At 14 days after grafting, the sections from control group revealed an inflammatory process which slightly decreases in intensity. The influx of leukocytes was maintained. A discreet fibroblasts differentiation, connective fibers and blood vessels neogenesis were observed in hypoderm. Animals tissues grafted with BC-\(\beta\)-keratose developed an inflammatory process with lower intensity. The healing process is represented by important connective hyperplasia and neogenesis of blood vessels, with the formation of prominent vascular-conjunctive buds. At lot BC-\(\beta\)-keratose with cells, the inflammatory process is discreet; the regeneration is markedly, represented by vascular, connective buds, and fibroblast proliferation. Connective fibers and capillary neogenesis are important. Figure 8: (a) illustrates the appearance of the two burned wounds A and X both covered with a dressing; (b) one illustrated the case of the burned wounds already grafted as one mentioned above. (c) is the appearance of grafted wound after an evolution of 14 days from the grafting date. One notices the appearance of hairs grown in the area of the grafted wound compared to the dorsal area of the rabbit previously cut for clinical study. Figure 9: Microscopic images of the stained histological sections (trichrome Masson staining) made on the tissues at different times after grafting (7 days, 14 days, 21 days). At 21 days after grafting at the animals from control group the inflammatory process is considerably reduced and the skin regeneration is represented by young conjunctive and vascular buds. The buds are covered with a superficial condensed cellular layer, without a basal membrane, suggesting a future stratified young and plane epithelium cells. Animals grafted with BC-\(\beta\)-keratose indicated that the re-epithelialization process is more advanced.