Gel formulated with Bryophyllum pinnatum leaf extract promotes skin wound healing in vivo by increasing VEGF expression: A novel potential active ingredient for pharmaceuticals

Edilane Rodrigues Dantas Araújo; Jacinthia Beatriz Xavier-Santos; Valéria Costa da Silva; Juliana Bessa Figueiredo de Lima; Jade Schlamb; Matheus de Freitas Fernandes-Pedrosa; Arnóbio Antônio da Silva Júnior; Raimundo Fernandes de Araújo Júnior; Thirumurugan Rathinasabapathy; Marvin Moncada; Debora Esposito; Gerlane Coelho Bernardo Guerra; Silvana Maria Zucolotto

doi:10.3389/fphar.2022.1104705

Gel formulated with Bryophyllum pinnatum leaf extract promotes skin wound healing in vivo by increasing VEGF expression: A novel potential active ingredient for pharmaceuticals

Front Pharmacol. 2023 Jan 12:13:1104705. doi: 10.3389/fphar.2022.1104705. eCollection 2022.

Authors

Edilane Rodrigues Dantas Araújo^{1

2}, Jacinthia Beatriz Xavier-Santos^{3

4}, Valéria Costa da Silva^{2

5}, Juliana Bessa Figueiredo de Lima², Jade Schlamb⁶, Matheus de Freitas Fernandes-Pedrosa^{3

4

5}, Arnóbio Antônio da Silva Júnior^{1

3

4

5}, Raimundo Fernandes de Araújo Júnior^{1

7}, Thirumurugan Rathinasabapathy⁶, Marvin Moncada^{6

8}, Debora Esposito^{6

9}, Gerlane Coelho Bernardo Guerra^{3

10}, Silvana Maria Zucolotto^{1

2

3

5

6}

Affiliations

¹ Postgraduate Program in Health Science, Federal University of Rio Grande do Norte (UFRN), Natal, Brazil.
² Department of Pharmacy, Research Group on Bioactive Natural Products, Federal University of Rio Grande do Norte, Natal, Brazil.
³ Postgraduate Program in Pharmaceutical Science, Federal University of Rio Grande do Norte, Natal, Brazil.
⁴ Biotechnology and Technology Laboratory, Department of Pharmacy, Federal University of Rio Grande do Norte, Natal, Brazil.
⁵ Postgraduate Program in Drug Development and Technological Innovation, Federal University of Rio Grande do Norte, Natal, Brazil.
⁶ Plants for Human Health Institute, North Carolina State University, Kannapolis, NC, United States.
⁷ Cancer and Inflammation Research Laboratory, Morphology Department, Biosciences Center, Federal University of Rio Grande do Norte, Natal, Brazil.
⁸ Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, United States.
⁹ Department of Animal Science, NC State University, Raleigh, NC, United States.
¹⁰ Department of Biophysics and Pharmacology, Biosciences Center, Federal University of Rio Grande do Norte, Natal, Brazil.

Abstract

Bryophyllum pinnatum (Crassulaceae) is used in traditional medicine for treating skin wounds. In our previous study, a topical gel containing B. pinnatum aqueous leaf extract showed a preclinical anti-inflammatory effect in in vivo acute edema models. In continuation, the present study aims to evaluate the phytochemical content and the stability of a formulation in gel containing B. pinnatum aqueous leaf extract and its healing properties and mechanism of action through an experimental model of induction of skin wounds in rats and in vitro assays. The animals were treated topically for 7 or 14 days with a formulation in gel containing extract at 5% or a placebo or Fibrinase^® in cream. In addition, to establish some quality control parameters, the total phenolic content (TPC), total flavonoid content (TFC), and a study focusing on the phytochemical and biological stability of a gel for 30 days at two different conditions (room temperature and 40°C/75% RH) were performed. Gel formulation containing extract showed a TPC and TFC of 2.77 ± 0.06 mg of gallic acid/g and 1.58 ± 0.03 mg of quercetin/g, respectively. Regarding the stability study, the formulation in gel showed no significant change in the following parameters: pH, water activity, chromatographic profile, and the content of the major compound identified in the extract. The gel formulation containing extract stimulated skin wound healing while reducing the wound area, as well as decreasing the inflammatory infiltrate, reducing the levels of IL-1β and TNF-α, and stimulating angiogenesis with increased expression of VEGF, an effect similar to Fibrinase. In conclusion, the gel formulation containing extract exhibited relevant skin wound healing properties and, therefore, has the potential to be applied as a novel active ingredient for developing wound healing pharmaceuticals.

Keywords: Crassulaceae; analytical marker; phenolic compounds; stability study; topical gel formulation; wound healing.

Grants and funding

This research had financial support by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior 630 (CAPES) finance code 001. This research was part of the Ph.D. thesis of ED of the Programa de Pós-Graduação em Ciências da Saúde at Federal University of Rio Grande do Norte. The in vitro analyses at Esposito D. were supported by North Carolina State University—Hatch Project #1016019 (NC02671) from the United States Department of Agriculture (USDA), National Institute of Food and Agriculture. Another part of work was supported by NCSU faculty start-up funds (MM 411050-02834) by MM. SZ is a research productivity fellow of the CNPq (Grant no. 313727/2020–1) and she was awarded with a Capes Print scholarship as Visiting Professor at Plants for Human Health Institute, North Carolina State University, United States.