## 3 Results and Discussion
### Histological Observations  
Histological analysis of wounds at day ten was performed by H&E and MT staining. In Figure 3, staining revealed the formation of thin epidermal layers in all samples. Furthermore, the application of CAPH-1D11-50 and CAPH-1D11-100 produced epidermal layers with irregular appearances compared to other groups. Previous studies have shown that alginate based wound dressings can promote fibroblast proliferation and collagen  
Figure 2: Wound contraction and assessment in response to application of Ca-alginate-PEGMA hydrogels with and without pan-specific anti-TGF-\(\beta\) mouse monoclonal antibody 1D11.16.8, evaluated over ten days. (**a**) Representative photographs of excisional wounds before and after application of Ca-alginate-PEGMA hydrogel (CAPH) loaded without or with 10, 50, and 100 \(\upmu\)g anti-TGF-\(\beta\) antibody (CAPH-1D11-10, CAPH-1D11-50, and CAPH-1D11-100, respectively), evaluated over 10 days. (**b**) Comparison of wound size over 10 days with initial wound size being 2 mm for all groups. Graphic presented as the mean of wound diameter size in mm \(\pm\) SEM, n = 5/group. (**c**) Epithelization. (**d**) Inflammation score of mice treated with CAPH with and without anti-TGF-\(\beta\) antibody. A four-point score was used (with 0 score (none), 1 score (scant), 2 score (moderate), and 3 score (abundant)), and is presented as a mean \(\pm\) SEM. n = 5/group. Significance is denoted as * \(p<0.05\) and ** \(p<0.01\). deposition [35; 50; 51]. In this study, the application of CAPH combined with a low anti-TGF-\(\beta\) dose, such as CAPH-1D11-10, improved wound epithelialization (evidenced by a tighter epidermis), but without loss of collagen deposition similar to CAPH. However, experimental observations should be performed for longer than ten days to determine if CAPH-1D11-10 could prevent scarring. In fibroblasts, the main promoter of collagen synthesis is TGF-\(\beta\)[52]. In Figure 3, MT staining of collagen fibers (in blue) revealed advanced collagen synthesis and remodeling with color intensity proportional to levels of deposited collagen [31; 53]. Wounds treated  
Figure 3: (**a**) Hematoxylin and eosin (H&E) and Masson’s trichrome (MT) stained microscopic sections at day 10. Two excisional wounds were created on each dorsal side before Ca-alignate-PEGMA hydrogels (CAPH), or CAPH loaded with 10, 50, and 100 ug anti-TGF-\(\beta\) antibody (CAPH-1D11-10, CAPH-1D11-50, and CAPH-1D11-100, respectively) were applied to the right side. The wound on the left side received only standard disinfection treatment (Control). Micrographs of stained tissues were captured under 100X magnification. Application of CAPH-1D11-10 produced a tighter epidermis than other treatments (black arrowheads), while treatment with CAPH-1D11-50 produced irregular epidermal layers. Application of CAPH-1D11-100 produced epidermis and collagen fragmentation (white arrowheads and black arrows, respectively). Edematous zones (asterisks) are also indicated in CAPH-1D11-50 and CAPH-1D11-100 treated wounds. (**b**) Quantitative analysis of collagen density from blue channel of MT-stained sections. CAPH-1D11-100 treated wounds exhibit the lowest collagen density, presented as a mean \(\pm\) SEM. n = 5/group. Significance is denoted as \({}^{*}p<0.05\) and \({}^{**}p<0.01\). with CAPH-1D11-100 showed reduced collagen deposition (Figure 3b) and fragmented epidermal layers. In wound healing, TGF-\(\beta\) not only promotes collagen synthesis, but is also responsible for preventing collagen degradation by regulating the synthesis of several ECM proteases and inducing the secretion of specific protease inhibitors [54; 55]. The appearance of fragmented collagen following the application of CAPH-1D11-50 and CAPH-1D11-100 could be attributed to enhanced collagen protease activity caused by TGF-\(\beta\) neutralization. However, the 1D11 antibody's affinity is biased towards TGF-\(\beta\)3 [56], the denominated \"scarless\" isoform of TGF-\(\beta\). [PERSON] et al.