Rodrigues Gabriela Marques, de Almeida Mara Elvira, Marcelino Sóstenes Apolo Correia, Fernandes Paula Bretas Ullmann, da Cruz Jessica Oliveira Pereira, Araújo Françoise Louanne, Ferreira Raquel da Silva, Botelho Ana Flávia Machado, Bedoya Francisco Javier, Cahuana Gladys Margot, Hitos Ana Belén, Soria Bernat, Costal-Oliveira Fernanda, Duarte Clara Guerra, Tejedo Juan R, Chávez-Olórtegui Carlos, Melo Marília Martins
Department of Veterinary Clinic and Surgery, Veterinary College, Federal University of Minas Gerais (UFMG), Belo Horizonte, MG, Brazil.
Department of Molecular Biology and Biochemical Engineering, Universidad Pablo de Olavide, Seville, Spain.
J Venom Anim Toxins Incl Trop Dis. 2024 Jul 22;30:e20240004. doi: 10.1590/1678-9199-JVATITD-2024-0004. eCollection 2024.
Loxoscelism refers to a set of clinical manifestations caused by the bite of spiders from the genus. The classic clinical symptoms are characterized by an intense inflammatory reaction at the bite site followed by local necrosis and can be classified as cutaneous loxoscelism. This cutaneous form presents difficult healing, and the proposed treatments are not specific or effective. This study aimed to evaluate the protective effect of mesenchymal stromal cells-derived secretome on dermonecrosis induced by spider venom in rabbits.
Sixteen rabbits were distributed into four groups (n = 4). Except for group 1 (G1), which received only PBS, the other three groups (G2, G3, and G4) were initially challenged with 10 μg of venom, diluted in 100 μL of NaCl 0.9%, by intradermic injection in the interscapular region. Thirty minutes after the challenge all groups were treated with secretome, except for group 2. Group 1 (G1-control group) received intradermal injection (ID) of 60 μg of secretome in 0.15 M PBS; Group 2 (G2) received 0.9% NaCl via ID; Group 3 (G3) received 60 μg of secretome, via ID and Group 4 (G4), received 60 μg of secretome by intravenous route. Rabbits were evaluated daily and after 15 days were euthanized, necropsied and skin samples around the necrotic lesions were collected for histological analysis.
Rabbits of G1 did not present edema, erythema, hemorrhagic halo, or necrosis. In animals from G2, G3, and G4, edema appeared after 6h. However, minor edema was observed in the animals of G2 and G3. Hemorrhagic halo was observed in animals, six hours and three days after, on G2, G3, and G4. Macroscopically, in G4, only one animal out of four had a lesion that evolved into a dermonecrotic wound. No changes were observed in the skin of the animals of G1, by microscopic evaluation. All animals challenged with venom showed similar alterations, such as necrosis and heterophilic infiltration. However, animals from G4 showed fibroblast activation, early development of connective tissue, neovascularization, and tissue re-epithelialization, indicating a more prominent healing process.
These results suggest that secretome from mesenchymal stromal cells cultured in a xeno-free and human component-free culture media can be promising to treat dermonecrosis caused after spiders bite envenoming.
洛矶山蜘蛛中毒是指由该属蜘蛛叮咬引起的一系列临床表现。典型的临床症状表现为叮咬部位强烈的炎症反应，随后出现局部坏死，可归类为皮肤型洛矶山蜘蛛中毒。这种皮肤型中毒伤口愈合困难，目前提出的治疗方法既不特异也无效。本研究旨在评估间充质基质细胞分泌组对兔蜘蛛毒液诱导的皮肤坏死的保护作用。
将16只兔子分为四组（每组n = 4）。除第1组（G1）仅接受磷酸盐缓冲液（PBS）外，其他三组（G2、G3和G4）最初在肩胛间区域皮内注射10μg毒液（用100μL 0.9%氯化钠稀释）进行攻击。攻击30分钟后，除第2组外，所有组均用分泌组进行治疗。第1组（G1-对照组）在0.15M PBS中皮内注射60μg分泌组；第2组（G2）皮内注射0.9%氯化钠；第3组（G3）皮内注射60μg分泌组；第4组（G4）静脉注射60μg分泌组。每天对兔子进行评估，15天后实施安乐死、剖检，并收集坏死病变周围的皮肤样本进行组织学分析。
G1组的兔子未出现水肿、红斑、出血晕或坏死。在G2、G3和G4组的动物中，6小时后出现水肿。然而，G2和G3组的动物仅观察到轻微水肿。在G2、G3和G4组的动物中，6小时和3天后观察到出血晕。宏观上，在G4组中，四只动物中只有一只出现了演变成皮肤坏死伤口的病变。通过显微镜评估，G1组动物的皮肤未观察到变化。所有接受毒液攻击的动物均表现出类似的改变，如坏死和嗜异性细胞浸润。然而，G4组的动物显示出成纤维细胞活化、结缔组织早期发育、新血管形成和组织再上皮化，表明愈合过程更为显著。
这些结果表明，在无血清和无人类成分的培养基中培养的间充质基质细胞分泌组有望用于治疗蜘蛛叮咬中毒后引起的皮肤坏死。
