# Clinical Efficacy of SPARC-Modified Mesenchymal Stem Cells for the Treatment of Dog Skin Wounds

## TL;DR

This study shows that modifying stem cells to produce more SPARC improves skin wound healing in dogs by boosting tissue regeneration and reducing inflammation.

## Contribution

The novel use of SPARC-overexpressing adipose-derived stem cells significantly enhances wound healing in dogs and human cell models.

## Key findings

- SPARC-modified stem cells improved skin regrowth, collagen production, and hair follicle regeneration in dog wounds.
- These cells also reduced inflammation and promoted new blood vessel formation.
- In human cell tests, SPARC-modified stem cells enhanced cell proliferation and wound healing processes.

## Abstract

Skin wounds are a common health issue in dogs, particularly for aging or diabetic animals, where healing can be challenging. Mesenchymal stem cells are promising for treatment but face limitations like poor survival at the wound site. This study explores a new strategy by using adipose-derived stem cells (ADSCs) that are engineered to produce high levels of a protein called SPARC. In a dog wound model, these modified cells significantly improved healing by enhancing skin regrowth, collagen production, new blood vessel formation, and hair follicle regeneration while reducing inflammation. Laboratory tests on human skin and blood vessel cells also showed that the secretions from these SPARC-enhanced stem cells promoted cell growth. Overall, combining SPARC with ADSCs presents a highly effective and promising approach for treating difficult skin wounds in dogs.

The number of pet dogs is increasing, and the number of working dogs (e.g., guide dogs, police dogs) is also gradually increasing. Skin wounds are a common clinical problem in dogs and tend to be more common in the clinic as mechanical wounds. The healing process of skin wounds is often influenced by a variety of factors, including infection, nutritional status, and immune response, while wound healing is more difficult in dogs with diabetes or aging dogs. Mesenchymal stem cells (MSCs) play an important role in skin healing and regeneration with their multidirectional differentiation potential and immunomodulatory function. However, the application of MSCs alone for the treatment of skin wounds may have certain limitations, such as low cell survival and a lack of localization. Therefore, it is important to find methods that can enhance the therapeutic effect of MSCs. Secreted protein acidic and rich in cysteine (SPARC), an extracellular matrix protein widely involved in regulating biological processes such as cell proliferation, migration, and matrix production, may enhance the efficacy of MSCs in skin wound healing. This study aims to systematically evaluate the therapeutic efficacy of SPARC-overexpressing adipose-derived mesenchymal stem cells (ADSCs) in promoting skin wound healing by establishing wound models in normal, diabetic, and aged mice and dogs, thereby validating their potential under diverse physiological and pathological conditions. For in vitro validation, we used hydrogen peroxide (H2O2) to induce Human Umbilical Vein Endothelial Cell (HUVEC) and Human Keratinocyte Cell (HaCaT) injury. All animals were randomly assigned to six experimental groups as follows: (1) Model group: Untreated wound (negative control); (2) HY group: Hydrogel alone (vehicle control); (3) Con group: Control-ADSCs (cell control); (4) Con-Exo&HY group: Control-ADSC exosomes in hydrogel; (5) SPARC group: oe-SPARC-ADSCs (treatment); (6) SPARC-Exo&HY group: oe-SPARC-ADSC exosomes in hydrogel (treatment). Separately, HUVEC and HaCaT cells were assigned to four experimental conditions: a blank control group, a model group, a control-ADSC-treated group, and an oe-SPARC-ADSC-treated group. ADSCs modified by SPARC significantly promoted re-epithelialization integrity, collagen deposition, inflammation reduction, angiogenesis, and hair follicle regeneration during wound healing in dog skin. HUVEC and HaCaT cells proliferated after adding oe-SPARC-ADSCs cell supernatant. Meanwhile, quantitative proteomic sequencing data analysis showed that SPARC could promote skin wound healing by enhancing cell adhesion, hyaluronic acid binding, and vascular smooth muscle contraction of ADSCs. Both in vitro cellular assays and in vivo wound-healing models suggest that the combination of SPARC and ADSCs for the treatment of skin wounds has broad application prospects.