Submit manuscript...
International Journal of
eISSN: 2573-2889

Molecular Biology: Open Access

Case Report Volume 4 Issue 6

Histologic evaluation of 5% Hyaluronic acid injection in oral tissue: A pilot study

KL Vandana, Shivani Singh, Liya Anil

Department of periodontics, College of Dental Sciences, India

Correspondence: Dr. Vandana KL, MDS, Senior Professor, Department of Periodontics, College of dental Sciences, Davangere– 577 004, Karnataka, India, Tel (08192) 231285, 231029, Fax 91-8192-251070

Received: December 02, 2019 | Published: December 26, 2019

Citation: Vandana KL, Singh S, Anil L. Histologic evaluation of 5% Hyaluronic acid injection in oral tissue: A pilot study. Int J Mol Biol Open Access.2019;4(6):206-208. DOI: 10.15406/ijmboa.2019.04.00122

Download PDF

Abstract

Background: Hyaluronic acid is a naturally derived, nonimmunogenic, nonadhesive glycosaminoglycan that plays a prominent role in various wound healing processes, as it as it is naturally angiogenic when degraded to small fragments.
Material and methodology: A case report was conducted on one male patient with operculum present bilaterally. Left side operculum was excised at baseline. The same day of excision, 5 % hyaluronic acid was injected on the right-side operculum which was excised on 15th day for post HA injection evaluation.
Results: Epithelium showed stratified squamous para-keratinization with focal epithelial hyperplasia entrapped with connective tissue. The connective tissue was moderately fibrous with focal areas of cellularity showing plum fibroblasts and fibrocytes.
Conclusion: Changes seen from pre to post HA injection are indicative of initial step towards increasing the size of a tissue clinically.

Keywords: hyaluronic acid, histology, epithelium, connective tissue

Introduction

Hyaluronic acid (HA) is a high molecular weight bio-polysaccharide, discovered in 1934, by Karl Meyer and his assistant, John Palmer in the vitreous of bovine eyes. Hyaluronic acid is a naturally occurring biopolymer, which has important biological functions in bacteria and higher animals including humans. It is found in most connective tissues and is particularly concentrated in synovial fluid, the vitreous fluid of the eye, umbilical cords and chicken combs. It is naturally synthesized by a class of integral membrane proteins called hyaluronan synthases and degraded by a family of enzymes called hyaluronidases.1

Hyaluronic acid (HA) plays a vital role in the synthesis of extracellular matrix molecules and epidermal cell interaction with the surrounding environment. It modulates cellular immunity by preventing infections and impeding allergic phenomena. One of its most important properties is that it can attach and hold large amounts of moisture; approximately 6L of water in just 1g. Hyaluronic acid is said to be an ultimate solution for moisture retention of the skin. Youthful skin is hydrated because it contains large amounts of HA in the dermis. However, as we age, the amount of HA in the skin decreases and by the time we become adults, this amount decreases to five percent of baseline.2

The challenging characteristic of conventional HAs in the use of topically applied anti-aging preparations has been that its molecules are 3,000nm in diameter, whereas the intercellular space is only 15 to 50nm and just 6 to 10nm at the hyaline membrane. This makes it impossible for conventionally produced HA to penetrate into deep layers of the dermis.3 There are several uses of hyaluronic acid in medical and dental field. In medical field it is used in facial rejuvenation,4 ophthalmology,5 osteoarthritis,6 bone formation,7 TMJ disorders,8 otolaryngology,9 cardiovascular disease,10 atrophic vaginitis,11 diabetic foot ulcer,12 peripheral nerve regeneration. 13 In dental field it is used in treatment of gingivitis,14 periodontis,15 gingival recession,16 in intrabony defects,17 oral lichen planus,18 recurrent aphthous ulceration,19 actinic cheilitis,20 implants,21 sinus lift, 22 in healing of extracted tooth sockets,23 through tissue engineering in gingival augmentation24 and dental pulp regeneration. 25

The use of hyaluronic acid in cosmetology for skin treatment is well known. However, the mechanism of action of HA after dermal injection is minimally explained. Recently, the use of HA injection for IDP deficiency treatment is in demand and there are few studies26–30 on its clinical effect. The skin anti-ageing treatment with hyaluronic acid is used effectively. However, the histologic evaluation of such ha injection in skin is minimally reported. Based on the dermal use of hyaluronic acid, successful clinical trial on 5% HA31 has been reported and prompted us to conduct the histologic evaluation in oral tissues to comprehend the mechanism of its action.

However, the histologic study of HA injection to understand its mechanism of action during IDP deficiency treatment has not been done so far. Hence, the present case-report was the first attempt to histologically evaluate the gingival tissue following 5% HA injection.

Methodology

In the current study attempt has been made to study the histological changes in oral tissue (operculum) 15 days after hyaluronic acid (HA) injection. A case report was conducted on one male patient with operculum present bilaterally. Left side operculum was excised at baseline. The same day of excision, 5 % hyaluronic acid was injected on the right side operculum which was excised on 15th day for post HA injection evaluation. Immediately after excision, tissue was transferred to 10% formalin and sent to oral pathology department for histologic examination. Staining was done with H&E stain and light microscope was used to evaluate the microscopic epithelium and connective tissue changes.

Results

Histo-pathological findings before Hyaluronic acid injection
Epithelium showed Stratified squamous para-keratinized with entrapped connective tissue. The connective tissue showed fibrous with few fibroblasts and fibrocytes interrupted with blood vessels. It also showed few lymphocytes, mast cells and neutrophils. Salivary acini, adipose tissues, nerve bundles and extravasated RBCs were also seen (Figure 1).

Figure 1 Histological picture at baseline.

Histo-pathological findings after Hyaluronic acid injection

Epithelium showed stratified squamous para-keratinization with focal epithelial hyperplasia entrapped with connective tissue. The connective tissue was moderately fibrous with focal areas of cellularity showing plum fibroblasts and fibrocytes with interspersed blood vessels. Inflammatory cells composed of lymphocytes, plasma cells, few neutrophils were seen. Areas of extravagated RBCs were also seen (Figure 2).

Figure 2 Histological picture after 15 days of post hyaluronic injection.

Discussion

Hyaluronic acid is a naturally derived, non-immunogenic, non-adhesive glycosaminoglycan that plays a prominent role in various wound healing processes, as it as it is naturally angiogenic when degraded to small fragments. Hyaluronic acid promotes early inflammation which is critical for initiating wound healing, but then moderates later stages of the process, allowing matrix stabilization and reduction of long-term inflammation. 32 Functions of HA include the following: hydration, lubrication of joints, a space filling capacity, and the framework through which cells migrate.33 The synthesis of HA increases during tissue injury and wound healing 34,35 and HA regulates several aspects of tissue repair, including activation of inflammatory cells to enhance immune response36 and the response to injury of fibroblasts37 and epithelial cells.38 HA also provides the framework for blood vessel formation 39 and fibroblast migration,40 that may be involved in tumor progression. The correlation of HA levels on the cell surface of cancer cells with the aggressiveness of tumors has also been reported.41

The histologic report of the current study suggests that the overall changes seen from pre to post HA injection were epithelial hyperplasia and moderately fibrous connective tissue with focal areas of fibroblast cellularity which are indicative of initial step towards increasing the size of a tissue clinically. The histologic HA effects were evaluated at 15 days in a single patient so that both pre and post HA injection tissue response can be appreciated from the same patient. The case report represents the first of the histologic effect of HA after gingival injection which serves as the basis for understanding the clinical voluminising effect of hyaluronic acid. The shortcoming of this pilot study was lack of patient's willingness to participate readily. Further histologic evaluation can be done to enhance the current study report using the similar methodological protocols in larger sample size participants.

Acknowledgments

None.

Ethical consent

None.

Conflicts of interest

The author declares there are no conflicts of interest.

References

  1. Necas J, Bartosikova L, Brauner P, et al. Hyaluronic acid (hyaluronan): a review. Veterinarni Medicina. 2008;53(8):397–411.
  2. Jegasothy SM, Zabolotniaia V, Bielfeldt S. Efficacy of a New Topical Nano-hyaluronic Acid in Humans. J Clin Aesthet Dermatol. 2014;7(3):27–29.
  3. Balaza EA, Band P. Hyaluronic acid: its structure and use. Cosmetic and Toiletries. 1984;99:65–72.
  4. Bogdan Allemann I, Baumann L. Hyaluronic acid gel (Juvéderm) preparations in the treatment of facial wrinkles and folds. Clin Interv Aging. 2008;3(4):629–634.
  5. Goa KL, Benfield P. Hyaluronic acid A review of its pharmacology and use as a surgical aid in ophthalmology, and its therapeutic potential in joint disease and wound healing. Drugs. 1994;47:536­–566.
  6. Pagnano M, Westrich G. Successful nonoperative management of chronic osteoarthritis pain of the knee: safety and efficacy of retreatment with intra­articular hyaluronans. Osteoarthritis Cartilage. 2005;13(9):751–761.
  7. Sasaki T, Watanabe C. Stimulation of osteoinduction in bone wound healing by high molecular hyaluronic acid. Bone. 1995;16(1):9–15.
  8. Basterzi Y, Sari A, Demirkan F, et al. Intraarticular hyaluronic acid injection for the treatment of reducing and nonreducing disc displacement of the temporomandibular joint. Ann Plast Surg. 2009;62(3):265–26­7.
  9. Angeli S. Hyaluronate gel stapedotomy. Otolaryngol Head Neck Surg. 2006;134(2):225–2­31.
  10. Francesca Bonafè, Marco Govoni, Emanuele Giordano, et al. Hyaluronan and cardiac regeneration. J Biomed Sci. 2014;21:100.
  11. Chen J, Geng L, Song X, et al. Evaluation of the efficacy and safety of hyaluronic acid vaginal gel to ease vaginal dryness: a multicenter, randomized, controlled, open­label, parallel­group, clinical trial. J Sex Med. 2013;10(6):1575–15­84.
  12. Chen CP, Hung W, Lin SH. Effectiveness of hyaluronic acid for treating diabetic foot: a systematic review and meta­analysis. Dermatologic Therapy. 2014;27:331–336.
  13. Seckel BR, Jones D, Hekimian KJ, et al. Hyaluronic acid through a new injectable nerve guide delivery system enhances peripheral nerve regeneration in the rat. J Neurosci Res. 1995;40(3):318–324.
  14. Sapna N, Vandana KL. Evaluation of hyaluronan gel (Gengigel) as a topical applicant in the treatment of gingivitis. J Investig Clin Dent. 2011;2(3):162­–170.
  15. Gontiya G, Galgali SR. Effect of hyaluronan on periodontitis: A clinical and histological study. J Indian Soc Periodontol. 2012;16(2):184–1­92.
  16. Fawzy E­ Sayed KM, Dahaba MA, Aboul ­Ela S, et al. Local application of hyaluronan gel in conjunction with periodontal surgery: a randomized controlled trial. Clin Oral Investig. 2012;16(4):1229–12­36.
  17. Vanden Bogaerde L. Treatment of infrabony periodontal defects with esterified hyaluronic acid: clinical report of 19 consecutive lesions. Int J Periodontics Restorative Dent. 2009;29(3):315–3­23.
  18. Nolan A, Badminton J, Maguire J, et al. The efficacy of topical hyaluronic acid in the management of oral lichen planus. J Oral Pathol Med. 2009;38(3):299–303.
  19. Nolan A, Baillie C, Badminton J, et al. The efficacy of topical hyaluronic acid in the management of recurrent aphthous ulceration. J Oral Pathol Med. 2006;35(8):461–465.
  20.  Lima Gs, Silva Gf, Gomes Ap, et al. Diclofenac In Hyaluronic Acid Gel: An Alternative Treatment For Actinic Cheilitis. J Appl Oral Sci. 2010;18(5):533–53­7.
  21. De Araújo Nobre M, Cintra N, Maló P. Peri­implant maintenance of immediate function implants: a pilot study comparing hyaluronic acid and chlorhexidine. Int J Dent Hyg. 2007;5(2):87–­94.
  22. Schwartz Z, Goldstein M, Raviv E, et al. Clinical evaluation of demineralized bone allograft in a hyaluronic acid carrier for sinus lift augmentation in humans: a computed tomography and histomorphometric study. Clin Oral Implants Res. 2007;18(2):204­–211.
  23. Mendes RM, Silva GA, Lima MF, et al. Sodium hyaluronate accelerates the healing process in tooth sockets of rats. Arch Oral Biol. 2008;53(12):1155­–1162.  
  24. Prato GP, Rotundo R, Magnani C, et al. An autologous cell hyaluronic acid graft technique for gingival augmentation: a case series. J Periodontol. 2003;74(2):262–26­7.
  25. Inuyama Y, Kitamura C, Nishihara T, et al. Effects of hyaluronic acid sponge as a scaffold on odontoblastic cell line and amputated dental pulp. J Biomed Mater Res B Appl Biomater. 2010;92(1):120–12­8.
  26. Becker W, Gabitov I, Stepanov M, et al. Minimally Invasive Treatment For Papillae Deficiencies In The Esthetic Zone: A Pilot Study. Clini Implant Dent and Related Res. 2010;12(1):1–­8.
  27. S Sadat Mansouri, M Ghasemi, Z Salmani, et al. Clinical application of hyaluronic acid gel for reconstruction of interdental papilla at the esthetic zone. Summer. 2013;25(3):152­–157.
  28. Awartani FA, Tatakis DN. Interdental papilla loss: treatment by hyaluronic acid gel injection: a case series. Clin Oral Investig. 2016;20(4):1775–17­80.
  29. Lee WP, Kim HJ, Yu SJ, et al. Six Month Clinical Evaluation of Interdental Papilla Reconstruction with Injectable Hyaluronic Acid Gel Using an Image Analysis System. J Esthet Restor Dent. 2016;28(4):221–2­30.
  30. Bertl K, Gotfredsen K, Jensen SS, et al. Can hyaluronan injections augment deficient papillae at implant-supported crowns in the anterior maxilla? A randomized controlled clinical trial with 6 months follow-up. Clin Oral Implants Res. 2017;28(9):1054–1061.
  31. Singh S, Vandana KL. Use of different concentrations of hyaluronic acid in interdental papillary deficiency treatment: a clinical study. J Indian Soc Periodontol. 2019;23(1):35–41.
  32. Saranraj P. Hyaluronic Acid Production and its Applications A Review. Int J Pharm Biol Arch. 2013;4(5):853–859.
  33. Toole BP. Hyaluronan: from extracellular glue to pericellular cue. Nat Rev Cancer. 2004;4(7):528–539.
  34. Juhlin L. Hyaluronan in skin. J Intern Med. 1997;242(1):61–66.
  35. Weigel PH, Fuller GM, LeBoeuf RD. A model for the role of hyaluronic acid and fibrin in the early events during the inflammatory response and wound healing. J Theor Biol. 1986;119(2):219–234.
  36. McKee CM, Penno MB, Cowman M, et al. Hyaluronan (HA) fragments induce chemokine gene expression in alveolar macrophages. The role of HA size and CD44. J Clin Invest. 1996;98(10):2403–2413.
  37. Bai KJ, Spicer AP, Mascarenhas MM, et al. The role of hyaluronan synthase 3 in ventilator-induced lung injury. Am J Respir Crit Care Med. 2005;172(1):92–98.
  38. Beck Schimmer B, Oertli B, Pasch T, et al. Hyaluronan induces monocyte chemoattractant protein-1 expression in renal tubular epithelial cells. J Am Soc Nephrol. 1998;9(12):2283–2290.
  39. Slevin M, Kumar S, Gaffney J. Angiogenic oligosaccharides of hyaluronan induce multiple signaling pathways affecting vascular endothelial cell mitogenic and wound healing responses. J Biol Chem. 2002;277(43):41046–14059.
  40. Li L, Heldin CH, Heldin P. Inhibition of platelet derived growth factor-BB-induced receptor activation and fibroblast migration by hyaluronan activation of CD44. J Biol Chem. 2006;281(36):26512–26519.
  41. Zhang L, Underhill CB, Chen L. Hyaluronan on the surface of tumor cells is correlated with metastatic behavior. Cancer Res. 1995;55(2):428–433.
Creative Commons Attribution License

©2019 Vandana, et al. This is an open access article distributed under the terms of the, which permits unrestricted use, distribution, and build upon your work non-commercially.