Enzymolysis behavior and safe collagenase management of porcine collagen fillers

Abstract

Collagen-based fillers are increasingly recognized as versatile biomaterials for soft-tissue augmentation, yet evidence guiding the enzymatic management of collagen-associated complications remains limited. Here, we performed a systematic characterization of two National Medical Products Administration-approved porcine type I collagen fillers (PCFs), glutaraldehyde cross-linked PCF (CL-PCF) and uncross-linked PCF (UCL-PCF), using hyaluronic acid (HA) filler as a clinical benchmark. CL-PCF exhibited superior mechanical robustness and markedly enhanced resistance to collagenase degradation relative to UCL-PCF. In a rabbit ear embolization model, both PCFs induced significantly milder vascular ischemic manifestations than HA. In vivo enzymolysis assessment in SD rats demonstrated that collagenase concentrations ≤ 300 IU/mL effectively facilitated the controlled degradation of PCFs without inducing notable tissue injury, whereas higher concentrations resulted in extensive matrix loss, hemorrhage, and focal skeletal muscle lysis with inflammatory infiltration. This study provides the first integrated mechanistic and safety-oriented framework for understanding the enzymolysis of collagen fillers and optimizing collagenase-based interventions. These findings offer clinical guidance for complication management and inform evidence-based decision-making in collagen filler applications.

Graphical Abstract

Keywords

Porcine collagen / Filler / Enzymolysis / Collagenase

References

[1] Salvatore L, Natali ML, Brunetti C, Sannino A, Gallo N. An update on the clinical efficacy and safety of collagen injectables for aesthetic and regenerative medicine applications. Polym (Basel), 2023, 15(4): 1020

[2] Kim JH, Kwon TR, Hong SW, Seok J, Kim JM, Hong JY, et al. . Comparative evaluation of the biodegradability and wrinkle reduction efficacy of Human-Derived collagen filler and hyaluronic acid filler. Aesthetic Plast Surg, 2019, 43(4): 1095-101

[3] Lee JH, Choi YS, Kim SM, Kim YJ, Rhie JW, Jun YJ. Efficacy and safety of Porcine collagen filler for nasolabial fold correction in asians: a prospective multicenter, 12 months follow-up study. J Korean Med Sci, 2014, 29(Suppl 3): S217-21

[4] Hirsch RJ, Narurkar V, Carruthers J. Management of injected hyaluronic acid induced Tyndall effects. Lasers Surg Med, 2006, 38(3): 202-4

[5] Li MY, Chien WY, Kang YN, Chen C. Efficacy and safety of fillers for the treatment of nasolabial folds: A network meta-Analysis of randomized controlled trials. Aesthetic Plast Surg, 2024, 48(17): 3452-62

[6] Tai HC, Liao YH, Chang YC, Yang CY, Horng SY, Kuo YS, et al. . An evaluation of skin and immunological responses after using a novel Cross-Linked Porcine-Based dermal injectable collagen with Lidocaine for nasolabial fold correction. J Clin Med, 2024, 13(17): 5241

[7] Chuang AD, Lin ET, Wu BQ, Lu ME, Chiang HM, Chu PN, et al. . Quantitative assessment of new frontiers in dermatochalasis and periorbital hyperpigmentation treatment: the role of Cross-Linked Porcine collagen. Aesthetic Plast Surg, 2025, 49(8): 2191-201

[8] Lehman A, Pilcher B, Roberts WE, Schlesinger TE, Vachon G. Postmarket experience of Polymethylmethacrylate-Collagen gel dermal filler. Dermatol Surg, 2020, 46(8): 1086-91

[9] Kroumpouzos G, Treacy P. Hyaluronidase for Dermal Filler Complications: Review of Applications and Dosage Recommendations. JMIR Dermatol. 202417;7:e50403.

[10] Smeraglia F, Del Buono A, Maffulli N. Collagenase clostridium histolyticum in dupuytren’s contracture: a systematic review. Br Med Bull, 2016, 118(1): 149-58

[11] Kaufman-Janette J, Joseph JH, Kaminer MS, Clark J, Fabi SG, Gold MH, et al. . Collagenase clostridium Histolyticum-aaes for the treatment of cellulite in women: results from two phase 3 Randomized, Placebo-Controlled trials. Dermatol Surg, 2021, 47(5): 649-56

[12] Li DJ, Berry CE, Wan DC, Longaker MT. Clinical, mechanistic, and therapeutic landscape of cutaneous fibrosis. Sci Transl Med, 2024, 16(766): eadn7871

[13] Castelanich D, Parra LA, Amado AM, Acevedo A, Velasquez L, Dicker V, et al. . Enzymatic management of facial overfilled syndrome: A case series and narrative review. J Cosmet Dermatol, 2025, 24(8): e70377

[14] Sun Y, Nie F, Wang G, Li Q, Xie H. An experimental study of the vascular embolism caused by Recombinant type III collagen implants and hyaluronic acid. J Cosmet Dermatol, 2023, 22(10): 2705-13

[15] Kim JH, Kwon TR, Lee SE, Jang YN, Han HS, Mun SK, et al. . Comparative evaluation of the effectiveness of novel hyaluronic Acid-Polynucleotide complex dermal filler. Sci Rep, 2020, 10(1): 5127

[16] Qiu H, Wang J, Hu H, Song L, Liu Z, Xu Y, et al. . Preparation of an injectable and photocurable carboxymethyl cellulose/hydroxyapatite composite and its application in cranial regeneration. Carbohydr Polym, 2024, 333: 121987

[17] Lorenc ZP, Nir E, Azachi M. Characterization of physical properties and histologic evaluation of injectable Dermicol-p35 porcine-collagen dermal filler. Plast Reconstr Surg, 2010, 125(6): 1805-13

[18] Wang M, Li W, Zhang Y, Tian W, Wang H. Comparison of Intra-arterial and subcutaneous testicular hyaluronidase injection treatments and the vascular complications of hyaluronic acid filler. Dermatol Surg, 2017, 43(2): 246-54

[19] Alnefaie RM, El-Sayed SA, Ramadan AA, Elmezien AI, El-Taher AM, Randhir TO, et al. . Physiological and anatomical responses of Faba bean plants infected with chocolate spot disease to chemical inducers. Life (Basel), 2023, 13(2): 392

[20] Sun Y, Jin M, Wang G, Xie H. Early warm compress treatment can promote recanalization of vascular embolisms and reduce tissue necrosis after polymethyl methacrylate injection. Sci Rep, 2023, 13(1): 1872

[21] Li X, Ji X, Chen K, Ullah MW, Yuan X, Lei Z, et al. . Development of finasteride/PHBV@polyvinyl alcohol/chitosan reservoir-type microspheres as a potential embolic agent: from in vitro evaluation to animal study. Biomater Sci, 2020, 8(10): 2797-813

[22] Zhao R, Qiu H, Liu S, Cao L, Yu D, Wang H. Quantifiable clinical efficacy of injectable Porcine collagen for the treatment of structural dark circles. J Cosmet Dermatol, 2021, 20(5): 1520-8

[23] Shen YH, Hsu CC. Triple cross-linked collagen, preparation method thereof, and cosmetic composition comprising the same. US patent application US20130039878A1. Published February 14, 2013.

[24] Pierre S, Liew S, Bernardin A. Basics of dermal filler rheology. Dermatol Surg, 2015, 41(Suppl 1): S120-6

[25] Choi MS. Basic rheology of dermal filler. Arch Plast Surg., Sundaram H, Voigts B, Beer K, Meland M. Comparison of the rheological properties of viscosity and elasticity in two categories of soft tissue fillers: calcium hydroxylapatite and hyaluronic acid. Dermatol Surg. 2010;36 Suppl 3:1859-65.

[26] Suesca E, Dias AMA, Braga MEM, de Sousa HC, Fontanilla MR. Multifactor analysis on the effect of collagen concentration, cross-linking and fiber/pore orientation on chemical, microstructural, mechanical and biological properties of collagen type I scaffolds. Mater Sci Eng C Mater Biol Appl, 2017, 77: 333-41

[27] Watt RP, Khatri H, Dibble ARG. Injectability as a function of viscosity and dosing materials for subcutaneous administration. Int J Pharm, 2019, 554: 376-86

[28] Lee W, Oh W, Oh SM, Yang EJ. Comparative effectiveness of different interventions of perivascular hyaluronidase. Plast Reconstr Surg, 2020, 145(4): 957-64

[29] Soares DJMD. Bridging a Century-Old problem: the pathophysiology and molecular mechanisms of HA Filler-Induced vascular occlusion (FIVO)-Implications for therapeutic interventions. Molecules, 2022, 27(17): 5398

[30] Nie F, Xie H, Wang G, An Y. Risk comparison of filler embolism between polymethyl methacrylate (PMMA) and hyaluronic acid (HA). Aesthetic Plast Surg, 2019, 43(3): 853-60

[31] Sito G, Manzoni V, Sommariva R. Vascular complications after facial filler injection: A literature review and Meta-analysis. J Clin Aesthet Dermatol, 2019, 12(6): E65-72

[32] Soares DJ, McCarthy AD. The impact of gel parameters on the dispersal and fragmentation of hyaluronic acid gel fillers within an artificial model of arterial embolism. Gels, 2024, 10(8): 530

[33] Nieswandt B, Watson SP. Platelet-collagen interaction: is GPVI the central receptor?. Blood, 2003, 102(2): 449-61

[34] Micheels P, Porcello A, Bezzola T, Perrenoud D, Quinodoz P, Kalia Y, et al. . Clinical perspectives on the injectability of Cross-Linked hyaluronic acid dermal fillers: A standardized methodology for commercial product benchmarking with Inter-Injector assessments. Gels, 2024, 10(2): 101

[35] Serwanja J, Wieland AC, Haubenhofer A, Brandstetter H, Schönauer E. A conserved strategy to attack collagen: the activator domain in bacterial collagenases unwinds triple-helical collagen. Proc Natl Acad Sci U S A, 2024, 121(16): e2321002121

[36] Ali KM, Huang Y, Amanah AY, Mahmood N, Suh TC, Gluck JM. Vitro biocompatibility and degradation analysis of Mass-Produced collagen fibers. Polym (Basel), 2022, 14(10): 2100

[37] AAlhayek A, Abdelsamie AS, Schönauer E, Camberlein V, Hutterer E, Posselt G, et al. . Discovery and characterization of synthesized and FDA-Approved inhibitors of clostridial and bacillary collagenases. J Med Chem, 2022, 65(19): 12933-55

[38] Demidova-Rice TN, Geevarghese A, Herman IM. Bioactive peptides derived from vascular endothelial cell extracellular matrices promote microvascular morphogenesis and wound healing in vitro. Wound Repair Regen, 2011, 19(1): 59-70

[39] Tian HW, Shi B, Chen JT, Gao Y, Han S, Hong W, et al. . Expert consensus on collagenase treatment of nodular complications after collagen (stimulant) implantation. China Med Cosmetology, 2024, 14(2): 1-7

[40] Soares DJ, Hynes SD, Yi CH, Shah-Desai S, Irving SC. Cosmetic Filler-Induced vascular occlusion: A rising threat presenting to emergency departments. Ann Emerg Med, 2024, 83(1): 59-67

[41] DeLorenzi C. New high dose pulsed hyaluronidase protocol for hyaluronic acid filler vascular adverse events. Aesthet Surg J, 2017, 37(7): 814-25

Funding

National Key Research and Development Program of China(2022YFC2409804)

Sichuan Provincial Natural Science Foundation(2025ZNSFSC1577)

Natural Science Foundation of Sichuan Province(2026NSFSC0532)

RIGHTS & PERMISSIONS

The Author(s)