Download evaluation of tolenorm oil and ointment in *near skin

Open Access Journal
Indian Journal of Medical Research and Pharmaceutical Sciences
January 2017; 4(1)
DOI: 10.5281/zenodo.232975
ISSN: ISSN: 2349-5340
Impact Factor: 3.052
EVALUATION OF TOLENORM OIL AND OINTMENT IN ‘NEAR SKIN’
MODEL FOR THE TREATMENT OF VITILIGO
Aruna.V*, Amruthavalli G.V, Satarupa Mukherjee, Gayathri Rajagopal
Dr.JRK’s Siddha Research and Pharmaceuticals Pvt., Ltd., No. 18 & 19 Perumal Koil Street, Kunrathur, Chennai600069
*
Abstract
Keywords:
Tolenorm oil, Tolenorm
ointment, Co-culture, Nearskin model, Vitiligo
treatment.
Combination of Tolenorm oil and Tolenorm ointment was tested on co-culture of
melanocytes and keratinocytes in vitro to understand whether the effect of these
Siddha drugs results in the transfer of melanin from melanocytes to keratinocytes.
The findings of the present study authenticates the established likely mechanism of
action of these Siddha drugs with reference to up-regulation of tyrosinase activity,
melanogenesis, dendrite formation and increase in dendrite length directly impact the
pigment transfer.
Introduction
Tolenorm oil and Tolenorm ointment are the proprietary Siddha medicines of Dr.JRK’s Siddha Research and
Pharmaceuticals. Extensive research work has been done on both these Siddha drugs and has established the likely
mechanism of action of these drugs at melanocyte level [1, 2].
Findings of our earlier cell culture studies have clearly shown that these siddha drugs may persuade the melanocytes
to produce tyrosinase enzyme and the process of melanogenesis [3]. Further the previous studies also have shown
the role of the above drugs in influencing the dendrite formation in melanocytes. The herbal ingredients used in the
formulation of Tolenorm oil and Tolenorm ointment have great Siddha legacy [4]. Further, these drugs also
haveproven usefulness in the treatment of Vitiligo ever since the drugs were introduced in the market.
All the above cell culture–based studies have undoubtedly established the clinical efficacy andthe likely mechanism
of action of these drugs. However, all the above cell culture studies were done at the level of melanocytes.
The mechanism of pigmentogenesis in mammalian system is quite complex [5]. It is not just the melanocytes are
responsible for the skin colour, but the ability of keratinocytes to receive the pigment effectively also plays a
significant role in the skin colour [6]. Any quantitative or qualitative defects in the melanocytes do affect in the
melanosome transfer to keratinocytes and that can result in pigmentory disorders [7].
Many AYUSH preparations are available in the market for the treatment of Vitiligo, however the real efficacy of all
such products remain mystery. For the effective treatment of Vitiligo, the drug must act both at melanocytes and
also in the transfer of melanosomes to keratinocytes.
In the present study, the efficacy of Tolenorm oil and Tolenorm ointment has been scientifically establishedthrough
the use ofco-culture of melanocytes and keratinocytes at the ratio at which they are present in the skin (near skin
model) [8, 9 & 10].
The biochemical responses like up-regulation of tyrosinase activity, melanogenesis and increase in the number and
length of dendrites due to the Siddha drugs whether also influencing in the transfer of melanin to keratinocyte can be
established systematically through ‘near skin model’ (co-culture method). Findings of the study are discussed in the
paper.
© Indian Journal of Medical Research and Pharmaceutical Sciences
http://www.ijmprs.com/
[6]
Open Access Journal
Indian Journal of Medical Research and Pharmaceutical Sciences
January 2017; 4(1)
DOI: 10.5281/zenodo.232975
ISSN: ISSN: 2349-5340
Impact Factor: 3.052
Materials and methods
Co-culture and treatment
B16F10 melanocytes and HaCaT cell keratinocytes were used for the co-culture experiment. HaCaT cells at 2.5
x10 5 per well in six-well plates were maintained in culture medium [8]. After two days, co-cultures of melanocytes
and keratinocytes were made in culture media (keratinocyte media + melanocyte media) with the initial seeding ratio
of keratinocytes to melanocytes being 5:1. After one day of co-culture, the combination of Tolenorm oil and
Tolenorm ointment at 40µg/ml (equal proportion) were prepared separately in double distilled water containing 5 %
glycerin and added into the co-culture medium and then were incubated for 48 hours. After 48 hours of incubation,
the cells were stained by Fontana – Masson stain [11] and were examined under microscope to evaluate the melanin
uptake by the keratinocytes as well as protrusion and elongation of dendrites.
Result
On microscopic examination, it was observed that nearly 40% of keratinocytes had received melanin. Further, the
dendrite protrusions from melanocytes leaning towards keratinocytes were also observed [Fig-1].
Fig-1 Co-culture experiment demonstrating dendrite formation and uptake of melanin by keratinocytes treated with
combination of Tolenorm oil and Tolenorm ointment.
Discussion
Findings of the ‘near skin model’ study have re-established the pre-existing clinical proofofTolenorm oil and
Tolenorm ointment for the treatment of Vitiligo. Theclinical proofwas gathered over the yearsfrom large number of
medical practitioners of Siddha, Ayurveda and Dermatology in different parts of India. Our previous studies have
clearly shown that both Tolenorm oil and Tolenorm ointment were effective in stimulating B16F10 melanocytes to
produce tyrosinase enzyme and the process of melanogenesis. Further, these Siddha drugs are also found to increase
the number and length of dendrites in melanocytes [12].
One of the active constituents of Piper nigrum thatpresent in Tolenorm oil has sufficient scientific proof for upregulating melanogenesis [13]. In our previous study, we have observed that Tolenorm oil and Tolenorm ointment
up-regulates melanogenesis both at pre- and post-transcriptional levels [14].
Findings of the present study have clearly shown that the combination of Tolenorm oil and Tolenorm ointment has
resulted in the effective transfer of melanin to keratinocytes as well.
The real cause of Vitiligo is idiopathic although a few predisposing factors were found to correlate with the disease.
One of the fundamental prerequisites for the treatment success is the number and quality of functional melanocytes
at the site or nearby the depigmented region.
© Indian Journal of Medical Research and Pharmaceutical Sciences
http://www.ijmprs.com/
[7]
Open Access Journal
Indian Journal of Medical Research and Pharmaceutical Sciences
January 2017; 4(1)
DOI: 10.5281/zenodo.232975
ISSN: ISSN: 2349-5340
Impact Factor: 3.052
Interestingly,several patients with functioning melanocytes were found to suffer from Vitiligo. The auto-immune
scavenging of the melanocytes is hypothesized for the above cause. The poor/ineffective transfer of the synthesized
melanin from melanosomes to keratinocytes is considered to be yet another reason for the above. There are several
biochemical and anatomical mechanisms are involved in melanin transfer. The formation of large number of
dendrites and their length play a major role in the melanin transfer.
The combination of Tolenorm oil and Tolenorm ointment seems to trigger cascading effect such as
1.
2.
3.
4.
Tyrosinase production
Melanin synthesis
Increase in the length and formation of dendrites in melanocytes
Transfer of melanin to keratinocytes
The question of why the combination of Tolenorm oil and Tolenorm ointment could not revolutionize the treatment
of Vitiligo lies more in the pathogenesis of Vitiligo than its therapeutics. All Vitiligo patients are not responding
evenly and equally to Tolenorm oil and Tolenorm ointment, whereas some are responding quickly and effectively.
We presume that the etiology of Vitiligo in all those who responds faster to Tolenorm oil and Tolenorm ointment
may likely to fall short of one or all of the above four defects. Perhaps, the etiology of Vitiligo in all those patients
who had responded poorly to Tolenorm oil and Tolenorm ointment might have fallen outside the premise of the
most known causes of Vitiligo.
A proper diagnosis for the real cause of Vitiligo is not available till date. Therefore, the patients are more often go
hopeless or get lost in the world of several products that come to the market with tall promises.
The co-culture findings highlight not just the science or efficacy of Tolenorm oil and Tolenorm ointment but indeed
about the great science of Siddha system of medicine. Above drugs are formulated fully in line with Siddha
principles using Siddha herbs. The study findings come not only as an assurance for the treatment of Vitiligo but
also validate great science of Siddha medicine of system.
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Open Access Journal
Indian Journal of Medical Research and Pharmaceutical Sciences
January 2017; 4(1)
DOI: 10.5281/zenodo.232975
ISSN: ISSN: 2349-5340
Impact Factor: 3.052
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