Vitamin D₃ and Dental Mesenchymal Stromal Cells

Round 1

Reviewer 1 Report

The topic of this article entitled “Vitamin D3 and dental mesenchymal stromal cells ” is aimed to summarize the current findings concerning the physiological effects of vitamin D3 on dental MSCs. This is an impacting and interesting article, with some interesting points of innovation. I ask authors to add some key concepts. - Authors report important concepts on tissue/organ regeneration. Something should also be reported on MSCs from dental and periapical tissues (Please, see and discuss: Tatullo, M.; Codispoti, B.; Pacifici, A.; Palmieri, F.; Marrelli, M.; Pacifici, L.; Paduano, F. Potential use of human periapical cyst-mesenchymal stem cells (hpcy-mscs) as a novel stem cell source for regenerative medicine applications. Front. Cell Dev. Biol 2017, 5, 103.) and report some specific concepts about translational applications of MSCs on humans (Please, see and discuss: Marrazzo, P.; Paduano, F.; Palmieri, F.; Marrelli, M.; Tatullo, M. Highly Efficient In Vitro Reparative Behaviour of Dental Pulp Stem Cells Cultured With Standardised Platelet Lysate Supplementation. Stem Cells Int 2016, 2016, 7230987.) - Authors should also give their point of view on nanotechnologies and nanomaterials on bone/dental procedures and their clinical applications (for example, discuss: Barry M, Pearce H, Cross L, Tatullo, M., Gaharwar AK. Advances in Nanotechnology for the Treatment of Osteoporosis. Curr. Osteoporos Rep 2016, 14, 87–94.) The authors must describe other type of tests to evaluate stress on different biomaterials, such as the three-points bending test, so to create some background to novel scaffolds in bone regeneration (for example, discuss: Marrelli, M.; Maletta, C.; Inchingolo, F.; Alfano, M.; Tatullo, M. Three-point bending tests of zirconia core/veneer ceramics for dental restorations. Int J Dent 2013, 2013, 831976.). - Conclusions should be improved with clear take-home messages. Minor suggestion: - In the whole text there are some typos here and there: authors should carefully revise the text before resubmission.

Author Response

REVIEWER COMMENT

The topic of this article entitled “Vitamin D3 and dental mesenchymal stromal cells ” is aimed to summarize the current findings concerning the physiological effects of vitamin D3 on dental MSCs. This is an impacting and interesting article, with some interesting points of innovation. I ask authors to add some key concepts. - Authors report important concepts on tissue/organ regeneration.

AUTHORS’ ANSWER

We are thankful for reviewing our manuscript. As recognized by the Reviewer, our paper aims to summarize the current information about the effect of different vitamin D3 forms in dental mesenchymal stromal cells. We have divided our review into several parts, in which we discuss the impact of vitamin D3 on the osteogenic differentiation, RANKL production and inflammatory response in dental MSCs. We intended not to focus on the common aspects of tissue regeneration, like scaffolds and growth factors, but rather on the physiological effects of vitamin D3. We sincerely recognize the importance of scaffolds and growth factors in the modern cell-based approaches.  These aspects were considered in the revised version (lines 398-409); however, we did not go into more detail to keep the readers focused.

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REVIEWER COMMENT

Something should also be reported on MSCs from dental and periapical tissues (Please, see and discuss: Tatullo, M.; Codispoti, B.; Pacifici, A.; Palmieri, F.; Marrelli, M.; Pacifici, L.; Paduano, F. Potential use of human periapical cyst-mesenchymal stem cells (hpcy-mscs) as a novel stem cell source for regenerative medicine applications. Front. Cell Dev. Biol 2017, 5, 103.)

AUTHORS’ ANSWER

The study of Tatullo et al. about MSCs like cells from the periapical cyst is mentioned in the revised version of the review, see Ref.12.

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REVIEWER COMMENT

and report some specific concepts about translational applications of MSCs on humans (Please, see and discuss: Marrazzo, P.; Paduano, F.; Palmieri, F.; Marrelli, M.; Tatullo, M. Highly Efficient In Vitro Reparative Behaviour of Dental Pulp Stem Cells Cultured With Standardised Platelet Lysate Supplementation. Stem Cells Int 2016, 2016, 7230987.)

AUTHORS’ ANSWER

This study is cited and discussed in the revised version of the review, see Refs. 126.

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REVIEWER COMMENT

Authors should also give their point of view on nanotechnologies and nanomaterials on bone/dental procedures and their clinical applications (for example, discuss: Barry M, Pearce H, Cross L, Tatullo, M., Gaharwar AK. Advances in Nanotechnology for the Treatment of Osteoporosis. Curr. Osteoporos Rep 2016, 14, 87–94.)

AUTHORS’ ANSWER

We have mentioned the importance of nanotechnologies in the revised version of the review and cite the study proposed by the Reviewer (Ref. 130). However, we avoided extensive discussion of this topic to remain in the scope of the review. We hope that the Reviewer understands our point.

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REVIEWER COMMENT

The authors must describe other type of tests to evaluate stress on different biomaterials, such as the three-points bending test, so to create some background to novel scaffolds in bone regeneration (for example, discuss: Marrelli, M.; Maletta, C.; Inchingolo, F.; Alfano, M.; Tatullo, M. Three-point bending tests of zirconia core/veneer ceramics for dental restorations. Int J Dent 2013, 2013, 831976.). –

AUTHORS’ ANSWER

We sincerely appreciate the three-point bending test to evaluate novel scaffolds. However, in our view, this method is not connected to the topic of vitamin D3 effects in dental MSCs. We will be thankful if Reviewer can provide some hints for such a connection.

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REVIEWER COMMENT

Conclusions should be improved with clear take-home messages. Minor suggestion: -

AUTHORS’ ANSWER

The conclusion section was added to the manuscript (see, lines 420-425)

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REVIEWER COMMENT

 In the whole text there are some typos here and there: authors should carefully revise the text before resubmission.

AUTHORS’ ANSWER

Our manuscript was corrected by a native speaker and all minor issues were edited.

 

Reviewer 2 Report

This article by Andrucov is focused on the role of Vitamin D3 on dental mesenchymal stromal cells. Dental tissues contain resident mesenchymal stromal cells (MSCs). The authors have reported that the existing literature suggests that dental MSCs possess the ability to convert vitamin D3 into the active 1,25(OH)2D3, which stimulates osteogenic differentiation and reduce the inflammatory response of dental MSCs. This interesting aspect can be used in dental procedures. More on vitamin D3 metabolism and its effects on biological structure must be added. The pros and cons of Vit. D3 use must be added. Other RCTs must be described. Report on stem cells from dental sources must be added. Report on the influence of extracellular matrix and scaffolds must be added.

Author Response

REVIEWER COMMENT

More on vitamin D3 metabolism and its effects on biological structure must be added.

AUTHORS’ ANSWER

The topic of vitamin D metabolism is described in detail: we provide the information on the systemic vitamin D metabolism (see, lines 67-80 and Figure 1); local vitamin D conversion in dental tissues (see, lines 90-132 and Figure 1); and vitamin D3 metabolite 24R,25(OH)2D3 (see, lines 286-311). As the effects of vitamin D3 in MSCs of other tissues are already extensively discussed in numerous other papers, we tried to focus on dental tissue derived MSCs. We hope that the Reviewer can understand our view.  If there are any aspects about the effects of vitamin D on dental MSCs that are not highlighted enough, we would be thankful if the Reviewer could provide us some hints.

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REVIEWER COMMENT

The pros and cons of Vit. D3 use must be added.

AUTHORS’ ANSWER

We are thankful for this important comment. This aspect is discussed in our revised version (see, lines 387-395). The following references were added concerning this question: Refs. 118-122

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REVIEWER COMMENT

Other RCTs must be described.

AUTHORS’ ANSWER

Unfortunately, to date, there are not so many randomized clinical trials on the application of vitamin D3 in dentistry. This fact is recognized in the revised version of our manuscript (see, lines393-395. We have added some RCT on vitamin D3 in other fields (Refs. 123, 124) and underlined the importance of further research in this direction (see, lines 393-395).

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REVIEWER COMMENT

Report on stem cells from dental sources must be added.

AUTHORS’ ANSWER

We have added a report about MSCs isolated from the periapical cyst; this point was also suggested by the other Reviewer (see Ref. 12). Besides, our review also mentions MSCs from other sources like periodontal ligaments, gingiva, dental pulp, dental follicle, apical papilla (see Refs. 6-11). To the best of our knowledge, we have mentioned all reports about the effect of vitamin D3 on the dental MSCs. If some study on the effect of vitamin D3 in dental MSCs is overseen, we would be deeply appreciating the hint from the Reviewer.

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REVIEWER COMMENT

Report on the influence of extracellular matrix and scaffolds must be added.

AUTHORS’ ANSWER

According to the suggestion of this Reviewer, as well as the recommendation of Reviewer 1, we discuss the importance of scaffolds and the potential effect of vitamin D3 on the integrin expression and the interaction of MSCs with the scaffolds in the revised version.

Reviewer 3 Report

Dear authors, congratulations on your review in this interesting field. This is a narrative review which thoughly analyses the effects of vit D3 on dental mesenchymal cells, which could help in future therapies. I enjoyed reading it and I believe it will be of interest for a broad range of readers. I have therefore recommended it for publication in this journal

Author Response

REVIEWER COMMENT

Dear authors, congratulations on your review in this interesting field. This is a narrative review which thoughly analyses the effects of vit D3 on dental mesenchymal cells, which could help in future therapies. I enjoyed reading it and I believe it will be of interest for a broad range of readers. I have therefore recommended it for publication in this journal

AUTHORS’ ANSWER

Thank you very much for the appreciation of our work. We have made some modifications to our Review according to the suggestions of other Reviewers. We hope that these changes will not change your positive evaluation of our manuscript.

Reviewer 4 Report

General comments:

This review article clearly states its aims and limitations. The abstract is clear and summarizes the subject. Further perspectives are reasonably  described.

However a sentence to conclude would be appropriate.

Particular comments and suggestions:

Please check your affiliations, no 2 does not belong to any of the authors...

Please number your chapters (sections).

line 35. (reviewed in [1]) is not appropriate. Only use a number for the reference.

You refer to previous published papers using the author's name or statements as: one study, another study, most studies etc. It would be better to use the same, for example the author's name or rephrase. Repeating over and over again: this study, recent study, few studies, some studies, our studies etc. is really disturbing.

line 111. One study reported that...Please rephrase one study, it is too vague.

line 117. Several studies investigated the effect of vitamin D3 metabolites on the expression of VDR in various dental MSCs. Please rephrase several studies and give references to this sentence.

line 133-140, 169-171, 207-209, 367-384. Please provide references.

line 189. Several studies investigated the effects of vitamin D3 on the RANKL and OPG production by 189 dental MSCs. Please rephrase several studies and give references to this sentence.

line 205 Several studies investigated the effect of vitamin 205 D3 on the inflammatory and/or immunomodulatory properties of dental MSCs. Same as above.

line 284 The biological role of 1,25(OH)2D3 has been intensively investigated and sufficiently clarified in numerous studies over the last 30-40 years. Same as above.

line 296 Few studies also showed biological effects of 24R,25(OH)2D3 in MSCs. Same as above.

line 359. A group of Prof. Civitelli (St. Louis University). Please refer as Civitelli et al (and collab) as you did in case of other authors.

Author Response

REVIEWER COMMENT

General comments:

This review article clearly states its aims and limitations. The abstract is clear and summarizes the subject. Further perspectives are reasonably  described.

However a sentence to conclude would be appropriate.

AUTHORS’ ANSWER

We are thankful for this Reviewer for the overall positive evaluation of our manuscript. Conclusion section was added to the review as suggested.

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REVIEWER COMMENT

Please check your affiliations, no 2 does not belong to any of the authors.

AUTHORS ANSWER

Christian Behm and Xiaohui Rausch-Fan belong to the affiliation 2; it is mentioned in the revised version.

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REVIEWER COMMENT

Please number your chapters (sections).

AUTHORS’ ANSWER

The chapters were numbered in the revised version according to Reviewers` suggestion.

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REVIEWER COMMENT

line 35. (reviewed in [1]) is not appropriate. Only use a number for the reference.

AUTHORS’ ANSWER.

This was corrected in the revised version.

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REVIEWER COMMENT

You refer to previous published papers using the author's name or statements as: one study, another study, most studies etc. It would be better to use the same, for example the author's name or rephrase. Repeating over and over again: this study, recent study, few studies, some studies, our studies etc. is really disturbing.

line 111. One study reported that...Please rephrase one study, it is too vague.

line 117. Several studies investigated the effect of vitamin D3 metabolites on the expression of VDR in various dental MSCs. Please rephrase several studies and give references to this sentence.

line 133-140, 169-171, 207-209, 367-384. Please provide references.

line 189. Several studies investigated the effects of vitamin D3 on the RANKL and OPG production by 189 dental MSCs. Please rephrase several studies and give references to this sentence.

line 205 Several studies investigated the effect of vitamin 205 D3 on the inflammatory and/or immunomodulatory properties of dental MSCs. Same as above.

line 284 The biological role of 1,25(OH)2D3 has been intensively investigated and sufficiently clarified in numerous studies over the last 30-40 years. Same as above.

line 296 Few studies also showed biological effects of 24R,25(OH)2D3 in MSCs. Same as above.

line 359. A group of Prof. Civitelli (St. Louis University). Please refer as Civitelli et al (and collab) as you did in case of other authors.

 

AUTHORS’ ANSWER.

All these points were considered in the revised version. Sentences were rephrased and References added as suggested by Reviewer.

Round 2

Reviewer 1 Report

Authors have revised properly the manuscript that now can be accepted

Reviewer 2 Report

The manuscript is acceptable in the current form