## 3 Results and Discussion
### Wound Contraction  
To determine if CAPH-1D11 could retain the viability of 1D11 and enable antibody release in situ, CAPH and CAPH-1D11 were assessed in a mouse model of dorsal bilateral excisional wound healing [38; 39; 40] to observe the impact of TGF-\(\beta\) neutralization on wound healing parameters. Wound closure was evaluated for ten days to ensure a comprehensive assessment of the impact of TGF-\(\beta\) neutralization on wound healing. During this period, it was assumed wound healing would progress through the fibroblast and keratinocyte proliferation phase and enter the first stage of matrix remodeling (on day seven), characterized by ECM formation and collagen deposition [11; 41]. Figure 2 illustrates the progress of wound closure over the course of the ten-day study. The findings show that the application of CAPH-1D11-10 resulted in the lowest wound diameter at day ten, while delays in wound closure were observed in response to CAPH-1D11-100. Overall, wound sizes did not vary significantly between groups at any experimental time point (Figure 2b). Expansion of the wound was observed in the first day and is often described in the early stages of excisional wound healing models as a result of tissue loosening [31; 42].  
Macroscopic assessment of the wounds showed no papule, exudation, hemorrhage, or necrosis, and granulation tissue was similar across all groups (data not shown). Macroscopic epithelialization and inflammation were also assessed (Figure 2c,d). TGF-\(\beta\) is a potent inhibitor of keratinocyte proliferation through the induction of G1 cell cycle arrest [43; 44]. After treatment with CAPH-1D11-10 and CAPH-1D11-50, epithelialization scores increased, coinciding with a reversal in the proliferative arrest of keratinocytes and promoting epidermal recovery. These findings agree with a study by [PERSON] et al. [45], where treatment with a TGF-\(\beta\) siRNA or crocodile oil accelerated re-epithelialization and reduced TGF-\(\beta\)1 and Smad3 protein and mRNA levels [46]. However, in the study presented here, epithelialization scores of wounds treated with CAPH-1D11-100 were similar to control wounds. This effect was possibly due to the negative impact of excessive TGF-\(\beta\) neutralization in the wound environment, resulting in non-healing chronic wounds where the ECM is not organized for efficient keratinization [47; 48]. This assumption may also explain the delay in wound closure observed following CAPH-1D11-100 treatment.  
In the inflammation phase of wound healing, TGF-\(\beta\) has dual roles as a pro-inflammatory stimulus early in the phase and as a potent anti-inflammatory chemoattractant of immune cells, such as neutrophils or monocytes, towards the end of this phase [49]. In Figure 2d, reduced inflammation scores in wounds following treatment with CAPH-1D11 may be explained by a decrease in immune cell infiltration through neutralization of the chemoattractant activity of TGF-\(\beta\).  
Figure 1: In vitro assessment of transforming growth factor-beta (TGF-\(\beta\)) neutralization by pan-specific anti-TGF-\(\beta\) mouse monoclonal antibody 1D11.16.8 released from loaded Ca-alginate-poly(ethylene glycol) methyl ether methacrylate (PEGMA) hydrogels. HEK-Blue TGF-\(\beta\) cells were used to evaluate the capacity of Ca-alginate-PEGMA hydrogels (CAPH) loaded with 1 and 5 \(\upmu\)g 1D11.16.8 (CAPH-1D1-1 and CAPH-1D11-5, respectively) to release stable antibodies and neutralize 20 pM rhTGF-\(\beta\)1. Dry-1D1 is a reconstituted antibody that was lyophilized and 1D11 is a soluble antibody without any drying. Each bar represents the mean \(\pm\) SEM of 3 independent assays conducted in triplicate. Significance denoted by \(\mathbf{{}^{\ast\ast}}\)_p_ < 0.01 and \(\mathbf{{}^{\ast\ast\ast}}\)_p_ < 0.0001.