Abstract
Backgroundhyaluronic acid (HA), a non-sulphated glycosaminoglycan, is present in synovial fluid, vitreous humour serum and many connective tissues. Pharmaceutical preparations of HA are used in clinical practice for wound healing, joint pain, kerato-conjunctivitis, asthma, mouth care, oesophageal-reflux, and gastritis. Moreover, it is used as a filler to counteract ageing and facial lipoatrophy. Our study aims at investigating the in vitro antiviral activity of a high molecular weight HA.Methodsthe MTT test was used to rule out the potential toxic effects of HA on the different cell lines used in the antiviral assays. The antiviral activity of HA against Coxsackievirus B5, Herpes Simplex Virus-1, Mumps Virus, Adenovirus-5, Influenza Virus A/H1N1, Human Herpesvirus-6, Porcine Parvovirus, Porcine Reproductive and Respiratory Syndrome Virus was assessed by virus yield assays.Resultsthe most effective inhibition was observed against Coxsackievirus B5, with 3Log reduction of the virus yield at 4 mg/ml, and a reduction of 3.5Log and 2Log, at 2 mg/ml and 1 mg/ml, respectively: the selectivity index was 16. Mumps virus was highly inhibited too showing a reduction of 1.7Log at 1 mg/ml and 1Log at 4 mg/ml and 2 mg/ml (selectivity index = 12). The selectivity index for Influenza Virus was 12 with the highest inhibition (1Log) observed at 4 mg/ml. Herpes Simplex Virus-1 and Porcine Parvovirus were mildly inhibited, whereas no antiviral activity was observed with respect to Adenovirus-5, Human Herpesvirus-6, Porcine Reproductive and Respiratory Syndrome Virus. No HA virucidal activity was ever observed against any of the viruses tested. Kinetic experiments showed that both Coxsackievirus B5 and Herpes simplex virus-1 replication were consistently inhibited, not influenced by the time of HA addition, during the virus replication cycle.Conclusionsthe spectrum of the antiviral activity exhibited by HA against both RNA and DNA viruses, known to have different structures (with or without envelope) and replication strategies, suggests a non specific mechanism of action, probably involving cell membrane-virus interaction steps. The results of the kinetic experiments support this hypothesis.
Highlights
Hyaluronic acid (HA) is a non-sulphated glycosaminoglycan which consists of alternately repeating D-glucuronic acid and N-acetylglucosamine units
In this study we investigated the in vitro effects of a high molecular weight HA against a wide group of viruses covering a large spectrum of structural features and replication strategies: ADV-5, Coxsackievirus B5 (COXB5), Herpes Simplex Virus type 1 (HSV-1), Human Herpesvirus-6 (HHV-6), Influenza Virus A/ H1N1, Mumps Virus (MV), Porcine Parvovirus (PPV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV)
Lack of Cytotoxicity Cell protection from lysis Initially, in order to rule out any direct cytotoxic effect of HA, dose-dependent experiments were performed exposing the five cell lines, employed in the experiments, to HA for different times (1 to 6 days, according to the protocols used for virus growth) followed by MTT assay
Summary
Hyaluronic acid (HA) is a non-sulphated glycosaminoglycan which consists of alternately repeating D-glucuronic acid and N-acetylglucosamine units. HA is commonly used for cosmetic interventions, as a filler to counteract ageing and facial lipoatrophy, especially in HIV patients [3]. The replication of Herpes Simplex Virus type 2 [4], Respiratory Syncytial virus [5] and retroviruses [6] is inhibited by HA, while the Adenovirus (ADV) one results enhanced [7]. Such limited and apparently controversial data demand further investigations in order to better understand the HA biological properties
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