In 2021, the chapel was declared part of the UNESCO World Heritage Site of 14th-century fresco cycles composed of 8 historical buildings in Padua city centre.

The space was where an open-air procession and sacred representation of the Annunciation to the Virgin had been played out for a generation before the chapel was built.

A number of 14th-century sources (Riccobaldo Ferrarese, Francesco da Barberino, 1312–1313) testify to Giotto's presence at the Arena Chapel's site.

The fresco cycle can be dated with a good approximation to a series of documentary testimonies: the purchase of the land took place on 6 February 1300; the bishop of Padua, Ottobono dei Razzi, authorised the building some time prior to 1302 (the date of his transferral to the Patriarcato of Aquileia); the chapel was first consecrated on 25 March 1303, the feast day of the Annunciation; on 1 March 1304 Pope Benedict XI granted an indulgence to whoever visited the chapel; one year later on 25 March 1305 the chapel received its definitive consecration.

In January 1305, friars from the nearby Church of the Eremitani filed a complaint to the bishop, protesting that Scrovegni had not respected the original agreement.

Scrovegni was transforming his private oratory into a church with a bell tower, thus producing unfair competition with the Eremitani's activities.

When one observes the lower frame of the triumphal arch, right above Saint Catherine of Alexandria's small altar piece, Giotto's perfect symmetry is altered by a fresco decoration representing two medallions with busts of female saints, a lunette with Christ in glory, and two episodes from the Passion (the prayer in the Garden of Gethsemane and the Flagellation of Christ, which together give an overall sense of disharmony.

The main focus of the unknown artist's work is constituted by six monumental scenes on the side walls of the chancel that depict the last period of Mary's earthly life.

The chapel was originally connected with the Scrovegni palace, which was built on what remained of the foundations of the elliptical ancient Roman arena.

In June 2001, following a preparation study lasting over 20 years, the Istituto Centrale per il Restauro (Central Institute for Restoration) of the Ministry for Cultural Activities, in collaboration with Padua's Town Hall in its capacity of owner of the Arena Chapel, started a full-scale restoration of Giotto's frescoes under the late Giuseppe Basile's technical direction.

Giuliano Pisani's studies argued that a number of commonly held beliefs concerning the chapel are groundless, among them, the notion that Dante inspired Giotto.

Pisani also argued against the conjecture that the Frati Gaudenti fraternity, of which Enrico Scrovegni was a member, influenced the content of Giotto's fresco cycle.

Pisani's arguments have not yet been widely embraced by the scholarly community, and that debates persist regarding the impetus for the chapel's creation and the reasons behind its design.

According to Pisani,[8] Giotto painted the chapel's inner surface following a comprehensive iconographic and decorative project devised by the Augustinian theologian, Friar Alberto da Padova.

Giuliano Pisani asserts that Giotto followed a careful and deliberate theological programme based on Saint Augustine and devised by Friar Alberto da Padova.

Avarice, far from being "absent" in Giotto's cycle, is portrayed with Envy, forming with it a fundamental component of a more comprehensive sin.

Allowing for this, the selection and iconography of the scenes is broadly comparable to other contemporary cycles; Giotto's innovation lies in the monumentality of his forms and the clarity of his compositions.

It starts from high up on the lunette of the triumphal arch, with the uncommon scene of God the Father instructing the Archangel Gabriel to perform the Annunciation to Mary.

The Vice-Virtue section of the Arena Chapel illustrates the philosophical-theological message underlying the overall project and is key to clarifying several points previously considered to be either obscure or the result of Giotto's only approximate theological knowledge.

Its cure (opposite wall) is Prudencia, Prudence, which in classical and theological terms is not "cautiousness" but "moral intelligence" or the capacity to distinguish good and evil.

According to Saint Augustine, Temperantia is the inner balance which ensures the will's stable dominion over instincts and keeps human desires within the boundaries of honesty.

Ethical virtue takes form in practical application, through action and behaviour that pertain both to the personal and the social sphere and affect human relations.

In order to be able to aspire to heavenly Paradise one needs divine teaching, the revelation of truth, with which one overcomes and transcends human reason, and to practice the theological virtues.

Only with the "medicine" of Charity (Karitas) can man overcome Selfishness and Envy (Invidia), which lead him to look with malevolent eyes (Latin in-vidēre) at his neighbour, who is also made by God in His likeness.

[citation needed] The sources of this extraordinary program were identified by Pisani in a number of passages of Saint Augustine's works.