Potential of Biofermentative Unsulfated Chondroitin and Hyaluronic Acid in Dermal Repair

Abstract

Chondroitin obtained through biotechnological processes (BC) shares similarities with both chondroitin sulfate (CS), due to the dimeric repetitive unit, and hyaluronic acid (HA), as it is unsulfated. In the framework of this experimental research, formulations containing BC with an average molecular size of about 35 KDa and high molecular weight HA (HHA) were characterized with respect to their rheological behavior, stability to enzymatic hydrolysis and they were evaluated in different skin damage models. The rheological characterization of the HHA/BC formulation revealed a G’ of 92 ± 3 Pa and a G” of 116 ± 5 Pa and supported an easy injectability even at a concentration of 40 mg/mL. HA/BC preserved the HHA fraction better than HHA alone. BTH was active on BC alone only at high concentration. Assays on scratched keratinocytes (HaCaT) monolayers showed that all the glycosaminoglycan formulations accelerated cell migration, with HA/BC fastening healing 2-fold compared to the control. In addition, in 2D HaCaT cultures, as well as in a 3D skin tissue model HHA/BC efficiently modulated mRNA and protein levels of different types of collagens and elastin remarking a functional tissue physiology. Finally, immortalized human fibroblasts were challenged with TNF-α to obtain an in vitro model of inflammation. Upon HHA/BC addition, secreted IL-6 level was lower and efficient ECM biosynthesis was re-established. Finally, co-cultures of HaCaT and melanocytes were established, showing the ability of HHA/BC to modulate melanin release, suggesting a possible effect of this specific formulation on the reduction of stretch marks. Overall, besides demonstrating the safety of BC, the present study highlights the potential beneficial effect of HHA/BC formulation in different damage dermal models.

Argan (Argania spinosa L.) Seed Oil Cake as a Potential Source of Protein-Based Film Matrix for Pharmaco-Cosmetic Applications

“…Immortalized human dermal fibroblasts (HDF-hTERT) were cultured in DMEM containing high glucose, L-glutamine, sodium bicarbonate, and sodium pyruvate, supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 100 μg/mL antifungal agent. The cells were growth on tissue culture plates (BD Bioscience-Falcon, San Jose, CA, USA) in a humidified atmosphere (95% air and 5% CO 2 , v/v ) at 37 °C, as previously reported [ 31 , 32 ] with slight modification to evaluate the specific effects of the ASPC-based films.…”

“…HDF-hTERT (2 × 10 4 cells/cm 2 ) were seeded on the films by two different methods: (a) the direct method in which different film amounts were added directly into the wells containing previously seeded cells; (b) an indirect method in which the cell medium was preliminarily conditioned for 24 h in the presence of the films and then added to the cells (30–40% cellular confluence in the 3-(4,5-dimethyltiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability test [ 32 ], or 100% confluence in the wound healing assay), at a theoretical final concentration ranging from 0.1 to 5.0 mg/mL for the MTT test and only 0.1 and 1 mg/mL for the wound healing assay. All experiments were run using film samples (1–2 mg) suspended in 1.0 mL of culture medium, and the reported amounts are those for the whole film and not the argan proteins contained in it.…”

Hyaluronic Acid-Based Injective Medical Devices: In Vitro Characterization of Novel Formulations Containing Biofermentative Unsulfated Chondroitin or Extractive Sulfated One with Cyclodextrins

“…Thermal stabilization obtained following the patented protocols releases different interactions/entanglements between GAG chains; these are not obtained for HHA + CS + cd. In fact, hybrid complexes can shield HHA from the attack of BTH, as already evaluated in the formulation based on HHA and BC for dermal applications [48]. However, it has to be noticed that the enzymatic degradation is definitely slower for the chemically modified hyaluronan counterparts.…”

Intradermal Injection of Hybrid Complexes of High- and Low-Molecular-Weight Hyaluronan: Where Do We Stand and Where Are We Headed in Regenerative Medicine?

“…Vestibular injections of HCCs were employed by Garavaglia et al to treat vulvovaginal atrophy, one of the symptoms of genitourinary syndrome, with a significant improvement in genital symptoms and sexual function, and high patient-reported satisfaction levels [ 82 ]. While HA–SC has mostly been studied as a medical device in the areas of orthopaedics, in vitro findings also point to the innovative formulation as a possible new therapy in several dermatological, cosmetic, and regenerative medicine applications [ 83 ]. Compared to HHC without chondroitin, the combined formula (HA–SC) works more efficiently for wound reparation, collagen, and elastin expression [ 84 ].…”